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Nikon biofilms
EFG1 is involved in filamentation and biofilm formation in C. tropicalis . (A) Formation of filamentous cells by wild-type and EFG1 deletion strains of C. tropicalis in the MYA3404 genetic background. Representative DIC microscopy images are shown for each time point. Scale bars are 5 μm. (B) Images of representative <t>biofilms</t> for EFG1 deletion mutants in C. albicans and C. trpicalis (two independent isolates) grown in wells of six-well polystyrene plates. (C) Biomass dry weights of the same strains shown in (B), grown in the same conditions. Error bars represent the SD of five replicates.
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Images

1) Product Images from "Finding a Missing Gene: EFG1 Regulates Morphogenesis in Candida tropicalis"

Article Title: Finding a Missing Gene: EFG1 Regulates Morphogenesis in Candida tropicalis

Journal: G3: Genes|Genomes|Genetics

doi: 10.1534/g3.115.017566

EFG1 is involved in filamentation and biofilm formation in C. tropicalis . (A) Formation of filamentous cells by wild-type and EFG1 deletion strains of C. tropicalis in the MYA3404 genetic background. Representative DIC microscopy images are shown for each time point. Scale bars are 5 μm. (B) Images of representative biofilms for EFG1 deletion mutants in C. albicans and C. trpicalis (two independent isolates) grown in wells of six-well polystyrene plates. (C) Biomass dry weights of the same strains shown in (B), grown in the same conditions. Error bars represent the SD of five replicates.
Figure Legend Snippet: EFG1 is involved in filamentation and biofilm formation in C. tropicalis . (A) Formation of filamentous cells by wild-type and EFG1 deletion strains of C. tropicalis in the MYA3404 genetic background. Representative DIC microscopy images are shown for each time point. Scale bars are 5 μm. (B) Images of representative biofilms for EFG1 deletion mutants in C. albicans and C. trpicalis (two independent isolates) grown in wells of six-well polystyrene plates. (C) Biomass dry weights of the same strains shown in (B), grown in the same conditions. Error bars represent the SD of five replicates.

Techniques Used: Microscopy

2) Product Images from "The LuxS/AI-2 Quorum-Sensing System of Streptococcus pneumoniae Is Required to Cause Disease, and to Regulate Virulence- and Metabolism-Related Genes in a Rat Model of Middle Ear Infection"

Article Title: The LuxS/AI-2 Quorum-Sensing System of Streptococcus pneumoniae Is Required to Cause Disease, and to Regulate Virulence- and Metabolism-Related Genes in a Rat Model of Middle Ear Infection

Journal: Frontiers in Cellular and Infection Microbiology

doi: 10.3389/fcimb.2018.00138

KEGG pathway analysis of genes downregulated in Streptococcus pneumoniae D39Δ luxS biofilms compared with the D39 wild-type biofilms.
Figure Legend Snippet: KEGG pathway analysis of genes downregulated in Streptococcus pneumoniae D39Δ luxS biofilms compared with the D39 wild-type biofilms.

Techniques Used:

In vitro biofilm growth of Streptococcus pneumoniae D39 wild-type and D39Δ luxS strains at different time-points after inoculation (6, 12, 18, and 24 h). The error bars are the standard deviation from the mean. Statistical significance was calculated using the Student's t -test, * p
Figure Legend Snippet: In vitro biofilm growth of Streptococcus pneumoniae D39 wild-type and D39Δ luxS strains at different time-points after inoculation (6, 12, 18, and 24 h). The error bars are the standard deviation from the mean. Statistical significance was calculated using the Student's t -test, * p

Techniques Used: In Vitro, Standard Deviation

Scanning electron microscopy (SEM) images of Streptococcus pneumoniae in vitro biofilms grown for 18 h. (A–C) are representative SEM images of the D39 wild-type strain. The wild-type strain biofilms were thick and organized with a significant depth. (D–F) are SEM images of the D39Δ luxS strain. The D39Δ luxS biofilms were thin and disorganized, and extracellular polymeric substance (EPS) was absent.
Figure Legend Snippet: Scanning electron microscopy (SEM) images of Streptococcus pneumoniae in vitro biofilms grown for 18 h. (A–C) are representative SEM images of the D39 wild-type strain. The wild-type strain biofilms were thick and organized with a significant depth. (D–F) are SEM images of the D39Δ luxS strain. The D39Δ luxS biofilms were thin and disorganized, and extracellular polymeric substance (EPS) was absent.

Techniques Used: Electron Microscopy, In Vitro

Confocal microscopy images of Streptococcus pneumoniae in vitro biofilms grown for 18 h. (A) Confocal microscopy image of the D39 wild-type strain biofilm. (B) Confocal microscopy image of the D39Δ luxS strain biofilm.
Figure Legend Snippet: Confocal microscopy images of Streptococcus pneumoniae in vitro biofilms grown for 18 h. (A) Confocal microscopy image of the D39 wild-type strain biofilm. (B) Confocal microscopy image of the D39Δ luxS strain biofilm.

Techniques Used: Confocal Microscopy, In Vitro

Scanning electron microscopy (SEM) images of rat bullae inoculated with Streptococcus pneumoniae D39 wild-type and D39Δ luxS . (A–C) are representative SEM images of rat bullae inoculated with the D39 wild-type strain. In rats colonized with the wild-type strain, dense biofilm/cell debris was deposited on the cilia, and the cilia were coagulated and completely covered with biofilm debris. (D–F) are representative SEM images of rat bullae inoculated with the D39Δ luxS strain. In rat bulla colonized with the D39Δ luxS strain, less biofilm debris was visible, although the cilia were coagulated. (G–I) are representative SEM images of rat bullae inoculated with medium (vehicle control). The vehicle control rat bulla were clean.
Figure Legend Snippet: Scanning electron microscopy (SEM) images of rat bullae inoculated with Streptococcus pneumoniae D39 wild-type and D39Δ luxS . (A–C) are representative SEM images of rat bullae inoculated with the D39 wild-type strain. In rats colonized with the wild-type strain, dense biofilm/cell debris was deposited on the cilia, and the cilia were coagulated and completely covered with biofilm debris. (D–F) are representative SEM images of rat bullae inoculated with the D39Δ luxS strain. In rat bulla colonized with the D39Δ luxS strain, less biofilm debris was visible, although the cilia were coagulated. (G–I) are representative SEM images of rat bullae inoculated with medium (vehicle control). The vehicle control rat bulla were clean.

Techniques Used: Electron Microscopy

Streptococcus pneumoniae D39 wild-type and D39Δ luxS planktonic and biofilm growth. (A) Planktonic growth optical density at 600 nm. (B) Quantification of biomass of in vitro biofilms grown for 18 h, using a CV-microplate assay. (C) Colony-forming unit (CFU) counts of in vitro biofilms grown for 18 h. Error bars are the standard deviation from the mean. Statistical significance was calculated using the Student's t -test, * p
Figure Legend Snippet: Streptococcus pneumoniae D39 wild-type and D39Δ luxS planktonic and biofilm growth. (A) Planktonic growth optical density at 600 nm. (B) Quantification of biomass of in vitro biofilms grown for 18 h, using a CV-microplate assay. (C) Colony-forming unit (CFU) counts of in vitro biofilms grown for 18 h. Error bars are the standard deviation from the mean. Statistical significance was calculated using the Student's t -test, * p

Techniques Used: In Vitro, Standard Deviation

3) Product Images from "Verticalization of bacterial biofilms"

Article Title: Verticalization of bacterial biofilms

Journal: Nature physics

doi: 10.1038/s41567-018-0170-4

Mechanics of cell reorientation in modeled biofilms, ( a-b ) Properties of individual cells at the time t r of reorientation, defined as the time of the peak of total force on the cell prior to it becoming vertical. Analyses are shown for all reorientation events among different biofilms simulated for a range of initial cell lengths ℓ 0 ( a ) Distributions of reorientation “surface pressure” p r , defined as the total contact force in the xy plane acting on a cell at time t r , normalized by the cell’s perimeter, versus cell cylinder length ℓ . The white dashed curve shows the average reorientation surface pressure ⟨ p r ⟩ as a function of ℓ . The magenta dashed curve shows the threshold surface pressure p t from linear stability analysis for a modeled cell under uniform pressure, depicted schematically in the inset, ( b ) Distributions of the logarithm of reorientation torque τ r , defined as the magnitude of the torque on a cell due to cell-cell contact forces in the z direction at time t r , for different cell cylinder lengths ℓ . The white dashed curve shows the average values ⟨log τ r ⟩ as a function of ℓ . The orange dashed curve shows the scaling τ t ~ ℓ 2 of the threshold torque for peeling from linear stability analysis for a modeled cell, depicted schematically in the inset, ( c ) Mean reorientation length ⟨ ℓ r ⟩ (red), defined as the average value of cell length at t r , and mean cell cylinder length ⟨ ℓ ⟩ (gray), defined as the average length of all horizontal cells over all times of biofilm growth, averaged over ten simulated biofilms, each with initial cell cylinder length ℓ 0 , plotted versus ℓ 0 . The inset shows the distribution of reorientation lengths (red) and horizontal surface-cell lengths (gray) for ℓ 0 = 1 μm. ( d ) Mean avalanche size ⟨ N ⟩, defined as the average size of a cluster of reorienting cells that are proximal in space and time ( Supplementary Figs. 8 - 10 ), versus initial cell length ℓ 0 for the experimental biofilm (red triangle) and the modeled biofilm (red circles). Open gray triangle and circles indicate the corresponding mean avalanche sizes for a null model. Inset shows a side view of cell configurations in the xy plane at times t r for all reorientation events in a simulated biofilm with ℓ 0 = 2.5 μm. Reorientation events are colored alike if they belong to the same avalanche. Scale bars: 10 μm and 1 hour.
Figure Legend Snippet: Mechanics of cell reorientation in modeled biofilms, ( a-b ) Properties of individual cells at the time t r of reorientation, defined as the time of the peak of total force on the cell prior to it becoming vertical. Analyses are shown for all reorientation events among different biofilms simulated for a range of initial cell lengths ℓ 0 ( a ) Distributions of reorientation “surface pressure” p r , defined as the total contact force in the xy plane acting on a cell at time t r , normalized by the cell’s perimeter, versus cell cylinder length ℓ . The white dashed curve shows the average reorientation surface pressure ⟨ p r ⟩ as a function of ℓ . The magenta dashed curve shows the threshold surface pressure p t from linear stability analysis for a modeled cell under uniform pressure, depicted schematically in the inset, ( b ) Distributions of the logarithm of reorientation torque τ r , defined as the magnitude of the torque on a cell due to cell-cell contact forces in the z direction at time t r , for different cell cylinder lengths ℓ . The white dashed curve shows the average values ⟨log τ r ⟩ as a function of ℓ . The orange dashed curve shows the scaling τ t ~ ℓ 2 of the threshold torque for peeling from linear stability analysis for a modeled cell, depicted schematically in the inset, ( c ) Mean reorientation length ⟨ ℓ r ⟩ (red), defined as the average value of cell length at t r , and mean cell cylinder length ⟨ ℓ ⟩ (gray), defined as the average length of all horizontal cells over all times of biofilm growth, averaged over ten simulated biofilms, each with initial cell cylinder length ℓ 0 , plotted versus ℓ 0 . The inset shows the distribution of reorientation lengths (red) and horizontal surface-cell lengths (gray) for ℓ 0 = 1 μm. ( d ) Mean avalanche size ⟨ N ⟩, defined as the average size of a cluster of reorienting cells that are proximal in space and time ( Supplementary Figs. 8 - 10 ), versus initial cell length ℓ 0 for the experimental biofilm (red triangle) and the modeled biofilm (red circles). Open gray triangle and circles indicate the corresponding mean avalanche sizes for a null model. Inset shows a side view of cell configurations in the xy plane at times t r for all reorientation events in a simulated biofilm with ℓ 0 = 2.5 μm. Reorientation events are colored alike if they belong to the same avalanche. Scale bars: 10 μm and 1 hour.

Techniques Used:

Development of experimental and modeled biofilms. ( a , b ) Top-down and perspective visualizations of the surface layer of ( a ) experimental and ( b ) modeled biofilms, showing positions and orientations of horizontal (blue) and vertical (red) surface-adhered cells as spherocylinders of radius R = 0.8 μm, with the surface shown at height z = 0 μm (brown). Cells with n z
Figure Legend Snippet: Development of experimental and modeled biofilms. ( a , b ) Top-down and perspective visualizations of the surface layer of ( a ) experimental and ( b ) modeled biofilms, showing positions and orientations of horizontal (blue) and vertical (red) surface-adhered cells as spherocylinders of radius R = 0.8 μm, with the surface shown at height z = 0 μm (brown). Cells with n z

Techniques Used:

Two-component fluid model for verticalizing cells in biofilms. ( a ) Schematic illustration of the two-component continuum model. Horizontal cells (blue) and vertical cells (red) are modeled, respectively, by densities ρ h and ρ v in two spatial dimensions. The total cell density ρ ~ tot is defined as ρ h + ξρ ν , where ξ is the ratio of vertical to horizontal cell footprints. ( b ) Radial densities ρ of vertical cells ( ρ v , red), horizontal cells ( ρ h , blue), and total density ( ρ ~ tot , black), versus shifted radial coordinate r ~ , defined as the radial position relative to the boundary between the mixed interior and the horizontal cell periphery. Results are shown for the continuum model (left; radial cell density in units of μm −2 ), the experimental biofilm (middle; radial cell density in each μm-sized bin averaged over an observation window of 50 minutes), and the agent-based model biofilm (right; radial cell density in each μm-sized bin averaged for ten biofilms over an observation window of 6 minutes). For the continuum model and the agent-based model biofilms the parameters were chosen to match those obtained from the experiment ( Supplementary Figs. 12 - 13 ). Inset in the left-most panel shows the fraction of vertical cells in the continuum model at a given radius from the biofilm center (gray regions contain no cells, color scale is the same as in Fig. 1 ).
Figure Legend Snippet: Two-component fluid model for verticalizing cells in biofilms. ( a ) Schematic illustration of the two-component continuum model. Horizontal cells (blue) and vertical cells (red) are modeled, respectively, by densities ρ h and ρ v in two spatial dimensions. The total cell density ρ ~ tot is defined as ρ h + ξρ ν , where ξ is the ratio of vertical to horizontal cell footprints. ( b ) Radial densities ρ of vertical cells ( ρ v , red), horizontal cells ( ρ h , blue), and total density ( ρ ~ tot , black), versus shifted radial coordinate r ~ , defined as the radial position relative to the boundary between the mixed interior and the horizontal cell periphery. Results are shown for the continuum model (left; radial cell density in units of μm −2 ), the experimental biofilm (middle; radial cell density in each μm-sized bin averaged over an observation window of 50 minutes), and the agent-based model biofilm (right; radial cell density in each μm-sized bin averaged for ten biofilms over an observation window of 6 minutes). For the continuum model and the agent-based model biofilms the parameters were chosen to match those obtained from the experiment ( Supplementary Figs. 12 - 13 ). Inset in the left-most panel shows the fraction of vertical cells in the continuum model at a given radius from the biofilm center (gray regions contain no cells, color scale is the same as in Fig. 1 ).

Techniques Used:

Global morphological properties of experimental and modeled biofilms, ( a ) Top-down (upper row) and side views (lower row) of experimental biofilms grown with 0.4 μg/mL A22 (magenta), without treatment (yellow), and with 4 μg/mL Cefalexin (cyan), following overnight growth (upper row) and 7 hours after inoculation (lower row). Scale bar: 10 μm. Insets show magnifications of 10 μm 2 -sized regions of top-down views taken from the peripheries of biofilms. ( b ) Expansion speed c *, defined as the speed of the biofilm edge along the surface, versus the initial cell cylinder length ℓ 0 for experimental biofilms (A22, magenta; no treatment, yellow; Cefalexin, cyan), agent-based model biofilms (black circles), and continuum model (dashed black curve). Expansion velocities were determined from a linear fit of the basal radius R B of the biofilm versus time, where R B is defined at each time point as the radius of a circle with area equal to that of the biofilm base. For experimental biofilms, the boundary was extracted from the normalized fluorescence data (see Methods for details). For each treatment, the vertical error bars show the standard error of the mean of the expansion speed and the horizontal error bars bound the measured initial cell cylinder length ( Supplementary Fig. 1 ). Inset: model cells with lengths and radii corresponding to the averages for different treatments, ( c ) Biofilm aspect ratio H/R B for experimental biofilms grown under different treatments, where the biofilm height is defined as H = 3 V ∕ 2 R B 2 , the height of a semi-ellipsoid with a circular base of radius R B and volume V equal to that of the biofilm. Error bars show the standard error of the mean. Inset: overlay of biofilm outlines from bottom row of panel ( a ). Color designations and treatments same as in panel ( a ).
Figure Legend Snippet: Global morphological properties of experimental and modeled biofilms, ( a ) Top-down (upper row) and side views (lower row) of experimental biofilms grown with 0.4 μg/mL A22 (magenta), without treatment (yellow), and with 4 μg/mL Cefalexin (cyan), following overnight growth (upper row) and 7 hours after inoculation (lower row). Scale bar: 10 μm. Insets show magnifications of 10 μm 2 -sized regions of top-down views taken from the peripheries of biofilms. ( b ) Expansion speed c *, defined as the speed of the biofilm edge along the surface, versus the initial cell cylinder length ℓ 0 for experimental biofilms (A22, magenta; no treatment, yellow; Cefalexin, cyan), agent-based model biofilms (black circles), and continuum model (dashed black curve). Expansion velocities were determined from a linear fit of the basal radius R B of the biofilm versus time, where R B is defined at each time point as the radius of a circle with area equal to that of the biofilm base. For experimental biofilms, the boundary was extracted from the normalized fluorescence data (see Methods for details). For each treatment, the vertical error bars show the standard error of the mean of the expansion speed and the horizontal error bars bound the measured initial cell cylinder length ( Supplementary Fig. 1 ). Inset: model cells with lengths and radii corresponding to the averages for different treatments, ( c ) Biofilm aspect ratio H/R B for experimental biofilms grown under different treatments, where the biofilm height is defined as H = 3 V ∕ 2 R B 2 , the height of a semi-ellipsoid with a circular base of radius R B and volume V equal to that of the biofilm. Error bars show the standard error of the mean. Inset: overlay of biofilm outlines from bottom row of panel ( a ). Color designations and treatments same as in panel ( a ).

Techniques Used: Fluorescence

4) Product Images from "Citrullination mediated by PPAD constrains biofilm formation in P. gingivalis strain 381"

Article Title: Citrullination mediated by PPAD constrains biofilm formation in P. gingivalis strain 381

Journal: NPJ Biofilms and Microbiomes

doi: 10.1038/s41522-019-0081-x

Δ8820 biofilms contain more gingipain-derived adhesin proteins than 381 biofilms. a Western blot analysis of biofilm lysates and planktonic cell lysates using an anti-adhesin primary antibody. Δ rgpA and Δ kgp planktonic cell lysates were used to determine the specific adhesin proteins (from top to bottom: Rgp44, Kgp39, Rgp27). Mfa1 was used as a loading control. All identified bands are shown. Samples derive from the same experiment and the blots were processed in parallel. b , c Quantification of b biofilm and c planktonic blots shown in a . ND, none detected d Suspended 381 and Δ8820 colony biofilms were treated with the anti-adhesin primary antibody and then images were acquired by immunogold labeling and SEM. Gingipain-derived adhesin proteins localized to the surface of 381 cells, the surface of Δ8820 cells, and the matrix around Δ8820 cells. Arrows indicate representative colloidal gold particles. SE secondary electron, BSE ackscatter electron. Scale bar: 2 µm
Figure Legend Snippet: Δ8820 biofilms contain more gingipain-derived adhesin proteins than 381 biofilms. a Western blot analysis of biofilm lysates and planktonic cell lysates using an anti-adhesin primary antibody. Δ rgpA and Δ kgp planktonic cell lysates were used to determine the specific adhesin proteins (from top to bottom: Rgp44, Kgp39, Rgp27). Mfa1 was used as a loading control. All identified bands are shown. Samples derive from the same experiment and the blots were processed in parallel. b , c Quantification of b biofilm and c planktonic blots shown in a . ND, none detected d Suspended 381 and Δ8820 colony biofilms were treated with the anti-adhesin primary antibody and then images were acquired by immunogold labeling and SEM. Gingipain-derived adhesin proteins localized to the surface of 381 cells, the surface of Δ8820 cells, and the matrix around Δ8820 cells. Arrows indicate representative colloidal gold particles. SE secondary electron, BSE ackscatter electron. Scale bar: 2 µm

Techniques Used: Derivative Assay, Western Blot, Labeling

Deletion of PPAD decreases secreted Rgp enzymatic activity and increases Rgp and Kgp enzymatic activity in biofilms. Rgp enzymatic activity in a planktonic cultures and c biofilm lysates, and Kgp enzymatic activity in b planktonic culture and d biofilm lysates was measured using a colorimetric assay. a Less Rgp activity was measured in the supernatants of Δ8820 cultures. b There were no differences in Kgp activity between 381 and Δ8820. Both c Rgp and d Kgp enzymatic activities were greater in Δ8820 biofilm lysates than 381 biofilm lysates. Data are representative of three independent experiments showing similar results ( n = 3). Error bars represent the standard deviation of technical replicates. The data were analyzed using the Student’s two-tailed t -test. * p
Figure Legend Snippet: Deletion of PPAD decreases secreted Rgp enzymatic activity and increases Rgp and Kgp enzymatic activity in biofilms. Rgp enzymatic activity in a planktonic cultures and c biofilm lysates, and Kgp enzymatic activity in b planktonic culture and d biofilm lysates was measured using a colorimetric assay. a Less Rgp activity was measured in the supernatants of Δ8820 cultures. b There were no differences in Kgp activity between 381 and Δ8820. Both c Rgp and d Kgp enzymatic activities were greater in Δ8820 biofilm lysates than 381 biofilm lysates. Data are representative of three independent experiments showing similar results ( n = 3). Error bars represent the standard deviation of technical replicates. The data were analyzed using the Student’s two-tailed t -test. * p

Techniques Used: Activity Assay, Colorimetric Assay, Standard Deviation, Two Tailed Test

Deletion of the gene encoding PPAD enhances biofilm formation. a 381 and Δ8820 biomass after 24 h was quantified by staining with safranin. b 24 h biomass of empty vector controls (381 pT-COW and Δ8820 pT-COW) and complemented Δ8820 (Δ8820 pT-C8820) was quantified by staining with safranin. Data in a and b are averages of three independent experiments ( n = 9). Error bars represent the standard deviation. The data in a were analyzed using the Student’s two-tailed t -test. The data in b were analyzed by ANOVA with Bonferroni post tests. * p
Figure Legend Snippet: Deletion of the gene encoding PPAD enhances biofilm formation. a 381 and Δ8820 biomass after 24 h was quantified by staining with safranin. b 24 h biomass of empty vector controls (381 pT-COW and Δ8820 pT-COW) and complemented Δ8820 (Δ8820 pT-C8820) was quantified by staining with safranin. Data in a and b are averages of three independent experiments ( n = 9). Error bars represent the standard deviation. The data in a were analyzed using the Student’s two-tailed t -test. The data in b were analyzed by ANOVA with Bonferroni post tests. * p

Techniques Used: Staining, Plasmid Preparation, Standard Deviation, Two Tailed Test

Δ8820 biofilms are comprised of more bacterial cells and protein than 381 biofilms. a 381 and Δ8820 were grown for 24 h on glass, stained with SYTO 9 (cells) and SYPRO Ruby (extracellular protein), and images were acquired by CLSM. Images are a top-down view of stacked maximum intensity projection images. Scale bar: 20 µm. b SYTO 9, c SYPRO Ruby, and d total biomass were quantified using Comstat2. Data are averages of two replicates ( n = 2). Error bars represent the standard deviation. The data were analyzed using the Student’s two-tailed t -test. * p
Figure Legend Snippet: Δ8820 biofilms are comprised of more bacterial cells and protein than 381 biofilms. a 381 and Δ8820 were grown for 24 h on glass, stained with SYTO 9 (cells) and SYPRO Ruby (extracellular protein), and images were acquired by CLSM. Images are a top-down view of stacked maximum intensity projection images. Scale bar: 20 µm. b SYTO 9, c SYPRO Ruby, and d total biomass were quantified using Comstat2. Data are averages of two replicates ( n = 2). Error bars represent the standard deviation. The data were analyzed using the Student’s two-tailed t -test. * p

Techniques Used: Staining, Confocal Laser Scanning Microscopy, Standard Deviation, Two Tailed Test

Deletion of the gene encoding PPAD results in increased matrix production. 381 and Δ8820 colony biofilms grown on plates were imaged by Cryo-SEM. Bacterial cells were visible on the surface and within 381 colony biofilms. The surface of Δ8820 colony biofilms were smooth and cells within the biofilms were coated or encased in a matrix-like substance. Scale bar: (top left) 10 µm, (top middle) 5 µm, (top right) 1 µm, (bottom left) 20 µm, (bottom middle) 20 µm, (bottom right) 2 µm
Figure Legend Snippet: Deletion of the gene encoding PPAD results in increased matrix production. 381 and Δ8820 colony biofilms grown on plates were imaged by Cryo-SEM. Bacterial cells were visible on the surface and within 381 colony biofilms. The surface of Δ8820 colony biofilms were smooth and cells within the biofilms were coated or encased in a matrix-like substance. Scale bar: (top left) 10 µm, (top middle) 5 µm, (top right) 1 µm, (bottom left) 20 µm, (bottom middle) 20 µm, (bottom right) 2 µm

Techniques Used:

Δ8820 cells produce a surface-associated extracellular substance that is not present on 381 cells. Bacterial cells from a 381 and b , c Δ8820 colony biofilms grown on plates were negatively stained with 0.5% aqueous uranyl acetate and then imaged by TEM. a 381 displayed typical cell morphology. Scale bar: (left) 2 µm and (right) 200 nm. b Δ8820 cells were (left) surrounded by and (right) coated with an extracellular substance that could be stained by uranyl acetate. Scale bar: (left) 2 µm and (right) 200 nm. c The extracellular substance in Δ8820 samples was capable of (left) forming a meshwork around cells and (right) appeared to extrude from specific sites at the cell surface. Scale bar: 200 nm
Figure Legend Snippet: Δ8820 cells produce a surface-associated extracellular substance that is not present on 381 cells. Bacterial cells from a 381 and b , c Δ8820 colony biofilms grown on plates were negatively stained with 0.5% aqueous uranyl acetate and then imaged by TEM. a 381 displayed typical cell morphology. Scale bar: (left) 2 µm and (right) 200 nm. b Δ8820 cells were (left) surrounded by and (right) coated with an extracellular substance that could be stained by uranyl acetate. Scale bar: (left) 2 µm and (right) 200 nm. c The extracellular substance in Δ8820 samples was capable of (left) forming a meshwork around cells and (right) appeared to extrude from specific sites at the cell surface. Scale bar: 200 nm

Techniques Used: Staining, Transmission Electron Microscopy

5) Product Images from "Synergistic Antifungal Effect of Amphotericin B-Loaded Poly(Lactic-Co-Glycolic Acid) Nanoparticles and Ultrasound against Candida albicans Biofilms"

Article Title: Synergistic Antifungal Effect of Amphotericin B-Loaded Poly(Lactic-Co-Glycolic Acid) Nanoparticles and Ultrasound against Candida albicans Biofilms

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.02022-18

Activity of biofilm (A) and biofilm biomass (B) are decreased significantly following treatment with a combination of ultrasound and AmB-NP compared with the control group. Ultrasonic irradiation parameters with a sound intensity of 0.30 W/cm 2 for 15 min were chosen, and the final concentration of drug employed in the AmB group and AmB-NP group was 4.0 μg/ml. The double asterisks denote a significant difference ( P
Figure Legend Snippet: Activity of biofilm (A) and biofilm biomass (B) are decreased significantly following treatment with a combination of ultrasound and AmB-NP compared with the control group. Ultrasonic irradiation parameters with a sound intensity of 0.30 W/cm 2 for 15 min were chosen, and the final concentration of drug employed in the AmB group and AmB-NP group was 4.0 μg/ml. The double asterisks denote a significant difference ( P

Techniques Used: Activity Assay, Irradiation, Concentration Assay

Scanning electron microscopy images of C. albicans biofilms on the surface of a rat subcutaneous catheter after 7-day treatments. The biofilm is attached to the catheter surface and has a complex structure with hyphae, yeast, and extracellular matrix in the control samples. The AmB-related treatments show deformed morphology of the biofilm, and treatment with ultrasound combined with AmB-NPs led to the complete inhibition of the biofilm, with cells remaining in the yeast form and being swollen and inactivated.
Figure Legend Snippet: Scanning electron microscopy images of C. albicans biofilms on the surface of a rat subcutaneous catheter after 7-day treatments. The biofilm is attached to the catheter surface and has a complex structure with hyphae, yeast, and extracellular matrix in the control samples. The AmB-related treatments show deformed morphology of the biofilm, and treatment with ultrasound combined with AmB-NPs led to the complete inhibition of the biofilm, with cells remaining in the yeast form and being swollen and inactivated.

Techniques Used: Electron Microscopy, Inhibition

Proteolytic (A) and phospholipase (B) enzymatic activities of C. albicans biofilms of different treatment groups. Significant reductions in proteinase and phospholipase enzyme activities were observed with the combination treatment with 4.0 μg/ml AmB. The double asterisks denote a statistically significant difference ( P
Figure Legend Snippet: Proteolytic (A) and phospholipase (B) enzymatic activities of C. albicans biofilms of different treatment groups. Significant reductions in proteinase and phospholipase enzyme activities were observed with the combination treatment with 4.0 μg/ml AmB. The double asterisks denote a statistically significant difference ( P

Techniques Used:

Morphological changes in catheter biofilm after seven consecutive days of treatment. C. albicans cells were stained with ConA and visualized by CLSM at a ×400 magnification. The control group of mature biofilms shows a dense structure on the catheter surface. In the group treated with ultrasound and AmB-NPs, only a single fungal colony remained on the catheter surface, and the biofilm was substantially eliminated. Bars, 50 μm.
Figure Legend Snippet: Morphological changes in catheter biofilm after seven consecutive days of treatment. C. albicans cells were stained with ConA and visualized by CLSM at a ×400 magnification. The control group of mature biofilms shows a dense structure on the catheter surface. In the group treated with ultrasound and AmB-NPs, only a single fungal colony remained on the catheter surface, and the biofilm was substantially eliminated. Bars, 50 μm.

Techniques Used: Staining, Confocal Laser Scanning Microscopy

6) Product Images from "Collective Vortex-Like Movement of Bacillus subtilis Facilitates the Generation of Floating Biofilms"

Article Title: Collective Vortex-Like Movement of Bacillus subtilis Facilitates the Generation of Floating Biofilms

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2018.00590

The effects of flagellar motility and chemotaxis on planktonic growth and floating biofilm formation. (A) Growth curves of wild-type and motility and chemotaxis mutants in shaking liquid cultures in biofilm inducing medium (MSgg). Error bars represent standard deviation. (B) Live cell counts of bacteria grown in standing liquid cultures before the floating biofilm is formed. Cultures were grown and harvested for CFU analyses (time post-inoculation is indicated in parenthesis). The differences between the wild-type and the indicated motility mutants were found as insignificant ( p > 0.5) in a two tailed paired student's t -test vs. the wild-type. (C) Images of pellicle formation process in wild-type and motility mutants in standing liquid cultures at 23°C.
Figure Legend Snippet: The effects of flagellar motility and chemotaxis on planktonic growth and floating biofilm formation. (A) Growth curves of wild-type and motility and chemotaxis mutants in shaking liquid cultures in biofilm inducing medium (MSgg). Error bars represent standard deviation. (B) Live cell counts of bacteria grown in standing liquid cultures before the floating biofilm is formed. Cultures were grown and harvested for CFU analyses (time post-inoculation is indicated in parenthesis). The differences between the wild-type and the indicated motility mutants were found as insignificant ( p > 0.5) in a two tailed paired student's t -test vs. the wild-type. (C) Images of pellicle formation process in wild-type and motility mutants in standing liquid cultures at 23°C.

Techniques Used: Chemotaxis Assay, Standard Deviation, Two Tailed Test

The impact of surface area on floating biofilm formation. Floating biofilm formation time of wild-type and motility and chemotaxis mutants in standing liquid cultures was monitored using a Nikon D800 camera (Nikon, Tokyo, Japan) and images were analyzed manually in order to identify time point of floating biofilm formation, for details refer to Materials and Methods. Error bars represent standard deviation. The effect of an increase of well diameter on the delay of pellicle formation was found insignificant within each given background ( P > 0.3). The Wild-type strain differed significantly from all motility mutants for each condition ( P
Figure Legend Snippet: The impact of surface area on floating biofilm formation. Floating biofilm formation time of wild-type and motility and chemotaxis mutants in standing liquid cultures was monitored using a Nikon D800 camera (Nikon, Tokyo, Japan) and images were analyzed manually in order to identify time point of floating biofilm formation, for details refer to Materials and Methods. Error bars represent standard deviation. The effect of an increase of well diameter on the delay of pellicle formation was found insignificant within each given background ( P > 0.3). The Wild-type strain differed significantly from all motility mutants for each condition ( P

Techniques Used: Chemotaxis Assay, Standard Deviation

The requirement for aggregation during floating biofilm formation . (A) Top-down images of the aggregates within a six well plate plates at 23°C after 20 h of incubation. (B) At either 10 h (v1), or 10 and 16 h (v2), and 10, 16, and 20 h (v3) post-inoculation the growth medium was collected, robustly vortexed and returned to the well. The delay in floating biofilm formation was the difference between un-vortexed controls within the same experiment, and vortexed samples. Conditioned media was harbored from WT (CM_WT) and luxS mutant (CM_Δ luxS ) floating biofilms grown under the same conditions and applied when indicated. Results are an average and standard deviation of 3 independent repeats performed in duplicates ( * P ≤ 0.05).
Figure Legend Snippet: The requirement for aggregation during floating biofilm formation . (A) Top-down images of the aggregates within a six well plate plates at 23°C after 20 h of incubation. (B) At either 10 h (v1), or 10 and 16 h (v2), and 10, 16, and 20 h (v3) post-inoculation the growth medium was collected, robustly vortexed and returned to the well. The delay in floating biofilm formation was the difference between un-vortexed controls within the same experiment, and vortexed samples. Conditioned media was harbored from WT (CM_WT) and luxS mutant (CM_Δ luxS ) floating biofilms grown under the same conditions and applied when indicated. Results are an average and standard deviation of 3 independent repeats performed in duplicates ( * P ≤ 0.05).

Techniques Used: Incubation, Mutagenesis, Standard Deviation

(A) The collective motion during floating biofilm formation. Motion * was defined as the mean of absolute difference between pixels of each two consecutive frames was calculated in time-lapse images series taken using a Nikon D800 camera of standing liquid cultures during floating biofilm formation, as described in Materials and Methods. (B) .
Figure Legend Snippet: (A) The collective motion during floating biofilm formation. Motion * was defined as the mean of absolute difference between pixels of each two consecutive frames was calculated in time-lapse images series taken using a Nikon D800 camera of standing liquid cultures during floating biofilm formation, as described in Materials and Methods. (B) .

Techniques Used:

The aggregation of EPS producers and motile cells during floating biofilm formation. Bacteria were grown in standing liquid cultures in six well plates at 23°C for 20 h. (A) Wild-type and cells harboring P tapA -mkateII and P hag -gfp ; or P tasA -mCherry and P hag -gfp . Cells were visualized by fluorescence microscopy; upper panel—phase contrast, lower panel—signal from GFP (green) and mKate/mCherry (red). The experiment shown here is a representative of 2 independent experiments; each done in at least duplicate. (B) Images of pellicle formation process in wild-type and the indicated mutants. WT-vor parental wild-type strain was collected at 20 h, robustly vortexed and returned to the well. (C) Enlargement of a well area defined with a white square in (B) Images are representative of three independent experiments, performed in triplicates. (D) Live cell counts of bacteria grown in standing liquid cultures before the pellicle is formed. Cultures were grown and harvested for CFU analyses 20 h post-inoculation. The differences between the wild-type and the indicated mutants were found as insignificant ( p > 0.5) in a two tailed paired student's t -test vs. the wild-type.
Figure Legend Snippet: The aggregation of EPS producers and motile cells during floating biofilm formation. Bacteria were grown in standing liquid cultures in six well plates at 23°C for 20 h. (A) Wild-type and cells harboring P tapA -mkateII and P hag -gfp ; or P tasA -mCherry and P hag -gfp . Cells were visualized by fluorescence microscopy; upper panel—phase contrast, lower panel—signal from GFP (green) and mKate/mCherry (red). The experiment shown here is a representative of 2 independent experiments; each done in at least duplicate. (B) Images of pellicle formation process in wild-type and the indicated mutants. WT-vor parental wild-type strain was collected at 20 h, robustly vortexed and returned to the well. (C) Enlargement of a well area defined with a white square in (B) Images are representative of three independent experiments, performed in triplicates. (D) Live cell counts of bacteria grown in standing liquid cultures before the pellicle is formed. Cultures were grown and harvested for CFU analyses 20 h post-inoculation. The differences between the wild-type and the indicated mutants were found as insignificant ( p > 0.5) in a two tailed paired student's t -test vs. the wild-type.

Techniques Used: Fluorescence, Microscopy, Two Tailed Test

7) Product Images from "Biological and immunotoxicity evaluation of antimicrobial peptide-loaded coatings using a layer-by-layer process on titanium"

Article Title: Biological and immunotoxicity evaluation of antimicrobial peptide-loaded coatings using a layer-by-layer process on titanium

Journal: Scientific Reports

doi: 10.1038/srep16336

Early biofilm formation (MOD) of S. aureus grown on the titanium plates with and without coating was tested. CS-(HA-AMPCol)7 and CS-(HA-AMPCol)10 exerted a significant inhibitory effect on biofilm formation (p
Figure Legend Snippet: Early biofilm formation (MOD) of S. aureus grown on the titanium plates with and without coating was tested. CS-(HA-AMPCol)7 and CS-(HA-AMPCol)10 exerted a significant inhibitory effect on biofilm formation (p

Techniques Used:

8) Product Images from "Experimental and Theoretical Investigation of Multispecies Oral Biofilm Resistance to Chlorhexidine Treatment"

Article Title: Experimental and Theoretical Investigation of Multispecies Oral Biofilm Resistance to Chlorhexidine Treatment

Journal: Scientific Reports

doi: 10.1038/srep27537

Scanning electron micrograph of a multispecies oral biofilm with mixed bacterial flora including numerous spirochetes. ( A ) three-week old biofilm after being treated with 2% CHX for 3 minutes is shown in ( A ); ( B–D ) higher magnification of ( A ) demonstrating tightly coiled spirochetes and a few damaged bacterial cells.
Figure Legend Snippet: Scanning electron micrograph of a multispecies oral biofilm with mixed bacterial flora including numerous spirochetes. ( A ) three-week old biofilm after being treated with 2% CHX for 3 minutes is shown in ( A ); ( B–D ) higher magnification of ( A ) demonstrating tightly coiled spirochetes and a few damaged bacterial cells.

Techniques Used:

Percentage of live cells in a multispecies biofilm over time. A 3-week-old biofilm was treated for 1, 3 and 10 minutes with the compounds indicated in the graph, respectively. The y-axis ratio corresponds to [live bacteria/(total bacteria)]. Two microliters of inactivator were added for 60 seconds immediately after each treatment. ( — • — No Treatment, CHX 1 minute with inactivator, CHX 1 minute without inactivator, CHX 3 min with inactivator, CHX 3 minutes without inactivator, CHX 10 min with inactivator, CHX 10 min without inactivator; CHX-Plus 1 minute with inactivator, CHX-Plus 1 minute without inactivator, CHX-Plus 3 minutes with inactivator, CHX-Plus 3 minutes without inactivator, CHX-Plus 10 minutes with inactivator, ○ CHX-Plus 10 minutes without inactivator).
Figure Legend Snippet: Percentage of live cells in a multispecies biofilm over time. A 3-week-old biofilm was treated for 1, 3 and 10 minutes with the compounds indicated in the graph, respectively. The y-axis ratio corresponds to [live bacteria/(total bacteria)]. Two microliters of inactivator were added for 60 seconds immediately after each treatment. ( — • — No Treatment, CHX 1 minute with inactivator, CHX 1 minute without inactivator, CHX 3 min with inactivator, CHX 3 minutes without inactivator, CHX 10 min with inactivator, CHX 10 min without inactivator; CHX-Plus 1 minute with inactivator, CHX-Plus 1 minute without inactivator, CHX-Plus 3 minutes with inactivator, CHX-Plus 3 minutes without inactivator, CHX-Plus 10 minutes with inactivator, ○ CHX-Plus 10 minutes without inactivator).

Techniques Used:

Scanning electron micrographs of a multispecies oral biofilm untreated and treated with CHX-Plus over time.
Figure Legend Snippet: Scanning electron micrographs of a multispecies oral biofilm untreated and treated with CHX-Plus over time.

Techniques Used:

Model prediction: the percentage of the live cell volume of a multispecies oral biofilm treated by CHX and the thickness of a naturally growing biofilm. The initial profile for each components are (0.056, 0.024, 0, 0, 1.0, 0, 0) for the volume fractions of susceptible, persister, live bacteria and EPS, as well as the concentrations for nutrient, antimicrobial agents and QS molecules, respectively. ( A ) Percentage of live bacterial cells in the biofilm at different time after being treated with CHX for 1, 3 and 10 minutes, respectively. The bottom curve corresponds to a controlled experiment where the biofilm is not treated with CHX (experimental data are obtained from 24 ); ( B ) Biofilm thickness of the controlled group. The experimental data are plotted with error bars.
Figure Legend Snippet: Model prediction: the percentage of the live cell volume of a multispecies oral biofilm treated by CHX and the thickness of a naturally growing biofilm. The initial profile for each components are (0.056, 0.024, 0, 0, 1.0, 0, 0) for the volume fractions of susceptible, persister, live bacteria and EPS, as well as the concentrations for nutrient, antimicrobial agents and QS molecules, respectively. ( A ) Percentage of live bacterial cells in the biofilm at different time after being treated with CHX for 1, 3 and 10 minutes, respectively. The bottom curve corresponds to a controlled experiment where the biofilm is not treated with CHX (experimental data are obtained from 24 ); ( B ) Biofilm thickness of the controlled group. The experimental data are plotted with error bars.

Techniques Used:

Bacteria in the multispecies oral biofilm after being treated with CHX up to 8 weeks. The initial profile for bacteria components: EPS and QS molecules are from the simulation in Fig. 1 at week 3. For the leftover concentrations of antimicrobial agents, they are fitted using the model as 1.67 × 10 −4 , 2.1 × 10 −4 and 3.75 × 10 −4 for CHX 1, 3, 10 minutes treatment correspondingly. The concentration of nutrient is given initially as 1. The percentage of live cell volume in the biofilm during recovery after treatment with CHX for 1, 3, 10 minutes and the control set (without treatment) are shown, respectively. The longer the biofilm is treated with CHX, the longer it takes for the bacterial cells to regain their populations (experimental data are obtained from 26 and Fig. 1 ).
Figure Legend Snippet: Bacteria in the multispecies oral biofilm after being treated with CHX up to 8 weeks. The initial profile for bacteria components: EPS and QS molecules are from the simulation in Fig. 1 at week 3. For the leftover concentrations of antimicrobial agents, they are fitted using the model as 1.67 × 10 −4 , 2.1 × 10 −4 and 3.75 × 10 −4 for CHX 1, 3, 10 minutes treatment correspondingly. The concentration of nutrient is given initially as 1. The percentage of live cell volume in the biofilm during recovery after treatment with CHX for 1, 3, 10 minutes and the control set (without treatment) are shown, respectively. The longer the biofilm is treated with CHX, the longer it takes for the bacterial cells to regain their populations (experimental data are obtained from 26 and Fig. 1 ).

Techniques Used: Concentration Assay

9) Product Images from "Vibrio cholerae Combines Individual and Collective Sensing to Trigger Biofilm Dispersal"

Article Title: Vibrio cholerae Combines Individual and Collective Sensing to Trigger Biofilm Dispersal

Journal: Current Biology

doi: 10.1016/j.cub.2017.09.041

RpoS- and HapR-Mediated Quorum Sensing Are Both Required for the Full Dispersal Response (A) Measurements of biofilm dispersal in mutants lacking rpoS , hapR , or both. The dispersal phenotype can be recovered in deletion mutants that carry chromosomal complementation constructs (n ≥ 24). (B) Expression of hapR , or rpoS , or both from an IPTG-inducible promoter in trans in a wild-type background causes biofilm dispersal (n ≥ 32). (C) Mutants deficient in various components of the V. cholerae quorum sensing circuit show attenuated or eliminated dispersal ability. Autoinducer synthases are cqsA (for CAI-1) and luxS (for AI-2), autoinducer receptors are cqsS (for CAI-1) and luxPQ (for AI-2), and strains carrying the luxO D47E or luxO D47A alleles are locked in the physiological “low cell density” or “high cell density” states, respectively [ 38 ] (n ≥ 12). Stars indicate a statistically significant difference from the WT (A and C) or empty vector control (B), with p
Figure Legend Snippet: RpoS- and HapR-Mediated Quorum Sensing Are Both Required for the Full Dispersal Response (A) Measurements of biofilm dispersal in mutants lacking rpoS , hapR , or both. The dispersal phenotype can be recovered in deletion mutants that carry chromosomal complementation constructs (n ≥ 24). (B) Expression of hapR , or rpoS , or both from an IPTG-inducible promoter in trans in a wild-type background causes biofilm dispersal (n ≥ 32). (C) Mutants deficient in various components of the V. cholerae quorum sensing circuit show attenuated or eliminated dispersal ability. Autoinducer synthases are cqsA (for CAI-1) and luxS (for AI-2), autoinducer receptors are cqsS (for CAI-1) and luxPQ (for AI-2), and strains carrying the luxO D47E or luxO D47A alleles are locked in the physiological “low cell density” or “high cell density” states, respectively [ 38 ] (n ≥ 12). Stars indicate a statistically significant difference from the WT (A and C) or empty vector control (B), with p

Techniques Used: Construct, Expressing, Plasmid Preparation

Integrating Nutrient and Autoinducer Sensing Yields Fine-Tuning of Biofilm Dispersal Decisions (A) By monitoring both nutrient supply and autoinducer concentration, V. cholerae cells can have four qualitatively different combinations of HapR and RpoS levels. This qualitative model predicts that a dispersal response is restricted to biofilms that are sufficiently large that a loss of flow or nutrient supply poses an immediate threat to cell viability. (B) Change in biofilm biomass as a function of biofilm size after glucose is removed from the flowing medium. (C) Change in biofilm biomass as a function of biofilm size after flow is halted. The biofilm size threshold for dispersal is accurately predicted by a mathematical model (red line), based on a calculation of the biofilm size that yields nutrient depletion after stopping the flow.
Figure Legend Snippet: Integrating Nutrient and Autoinducer Sensing Yields Fine-Tuning of Biofilm Dispersal Decisions (A) By monitoring both nutrient supply and autoinducer concentration, V. cholerae cells can have four qualitatively different combinations of HapR and RpoS levels. This qualitative model predicts that a dispersal response is restricted to biofilms that are sufficiently large that a loss of flow or nutrient supply poses an immediate threat to cell viability. (B) Change in biofilm biomass as a function of biofilm size after glucose is removed from the flowing medium. (C) Change in biofilm biomass as a function of biofilm size after flow is halted. The biofilm size threshold for dispersal is accurately predicted by a mathematical model (red line), based on a calculation of the biofilm size that yields nutrient depletion after stopping the flow.

Techniques Used: Concentration Assay, Flow Cytometry

Characterization of the V. cholerae Biofilm Dispersal Process (A) Biofilms grown at a flow rate 0.1 μL/min (until they reach a diameter of 20–25 μm) undergo a substantial change in biomass within 4 hr after the flow rate is stopped, reduced, or increased. Error bars indicate SD of n ≥ 24 replicas. See also Movie S1 . (B) Biomass change of biofilms for which a range of solutes were altered. Biofilms were grown to a diameter of 20–25 μm before composition of the growth medium was modified to either lack glucose, lack oxygen, or include 250 nM of nitric oxide for 4 hr. Control experiments refer to conditions in which the medium composition is unchanged and flow is kept constant (“keep flow”) but the tubing is exchanged (“keep flow † ”). Stars indicate a statistically significant difference from the “keep flow” condition (p
Figure Legend Snippet: Characterization of the V. cholerae Biofilm Dispersal Process (A) Biofilms grown at a flow rate 0.1 μL/min (until they reach a diameter of 20–25 μm) undergo a substantial change in biomass within 4 hr after the flow rate is stopped, reduced, or increased. Error bars indicate SD of n ≥ 24 replicas. See also Movie S1 . (B) Biomass change of biofilms for which a range of solutes were altered. Biofilms were grown to a diameter of 20–25 μm before composition of the growth medium was modified to either lack glucose, lack oxygen, or include 250 nM of nitric oxide for 4 hr. Control experiments refer to conditions in which the medium composition is unchanged and flow is kept constant (“keep flow”) but the tubing is exchanged (“keep flow † ”). Stars indicate a statistically significant difference from the “keep flow” condition (p

Techniques Used: Flow Cytometry, Modification

Induction of the General Stress Response and Quorum Sensing During Biofilm Dispersal (A) Averaged space-time heatmaps of RpoS levels during biofilm growth and dispersal, measured via an RpoS-mRuby3 translational fusion, before and after glucose removal, flow-stop, or the control condition (no change in flow or nutrient supply). All spatial coordinates in the biofilm are measured in terms of distance to the biofilm center of mass, d CM . See Figure S1 for how space-time heatmaps are generated and Figure S3 for data from a transcriptional rpoS reporter. Biofilms for each condition (n = 15) were binned and averaged in space and time, and statistically significant differences (p
Figure Legend Snippet: Induction of the General Stress Response and Quorum Sensing During Biofilm Dispersal (A) Averaged space-time heatmaps of RpoS levels during biofilm growth and dispersal, measured via an RpoS-mRuby3 translational fusion, before and after glucose removal, flow-stop, or the control condition (no change in flow or nutrient supply). All spatial coordinates in the biofilm are measured in terms of distance to the biofilm center of mass, d CM . See Figure S1 for how space-time heatmaps are generated and Figure S3 for data from a transcriptional rpoS reporter. Biofilms for each condition (n = 15) were binned and averaged in space and time, and statistically significant differences (p

Techniques Used: Flow Cytometry, Generated

10) Product Images from "The biofilm matrix destabilizers, EDTA and DNaseI, enhance the susceptibility of nontypeable Hemophilus influenzae biofilms to treatment with ampicillin and ciprofloxacin"

Article Title: The biofilm matrix destabilizers, EDTA and DNaseI, enhance the susceptibility of nontypeable Hemophilus influenzae biofilms to treatment with ampicillin and ciprofloxacin

Journal: MicrobiologyOpen

doi: 10.1002/mbo3.187

NTHi 502 static biofilm biomass and planktonic cell viability in the presence of divalent cations. Biofilms were formed in the presence of twofold increases in concentration of Mg + (A), Ba 2+ (B), Ca 2+ (C), Mn 2+ (D), Zn 2+ (E), and Fe 2+ (F) cations in CDM for 24 h at 37°C, 5% CO 2 . Circles represent biofilm biomass. Squares represent planktonic cell growth. Planktonic cell viability was measured by CFU counts and biofilm biomass quantitated with A595 measurement of crystal violet staining. Graphs show values relative to no added cation control. Dotted line indicates 100% of no added cation control values. In the presence of Fe 2+ , Mn 2+ , and Zn 2+ , biofilm formation and planktonic cell viability generally decreased as the cation concentration increased. Biofilms were increased in the presence of Ba 2+ and Ca 2+ in conjunction with an increase in planktonic cell viability. Mg 2+ was the only cation to result in increasing biofilm formation without an increase in planktonic cell viability. NTHi, nontypeable Hemophilus influenzae ; CDM, chemically defined medium pH 9.
Figure Legend Snippet: NTHi 502 static biofilm biomass and planktonic cell viability in the presence of divalent cations. Biofilms were formed in the presence of twofold increases in concentration of Mg + (A), Ba 2+ (B), Ca 2+ (C), Mn 2+ (D), Zn 2+ (E), and Fe 2+ (F) cations in CDM for 24 h at 37°C, 5% CO 2 . Circles represent biofilm biomass. Squares represent planktonic cell growth. Planktonic cell viability was measured by CFU counts and biofilm biomass quantitated with A595 measurement of crystal violet staining. Graphs show values relative to no added cation control. Dotted line indicates 100% of no added cation control values. In the presence of Fe 2+ , Mn 2+ , and Zn 2+ , biofilm formation and planktonic cell viability generally decreased as the cation concentration increased. Biofilms were increased in the presence of Ba 2+ and Ca 2+ in conjunction with an increase in planktonic cell viability. Mg 2+ was the only cation to result in increasing biofilm formation without an increase in planktonic cell viability. NTHi, nontypeable Hemophilus influenzae ; CDM, chemically defined medium pH 9.

Techniques Used: Concentration Assay, Staining

Effect of EDTA and DNaseI in combination with antibiotics on biofilm cell viability. Biofilms were formed in the flow cell model of NTHi biofilm development and treated with EDTA or DNaseI, alone or in combination with ampicillin or ciprofloxacin. Bacterial viability was assessed with live/dead staining kit and imaged using CLSM. Quantitative analysis of dead cell biomass was performed using COMSTAT of the proridium idodide channel (A). Data presented as mean ± SEM (*** P
Figure Legend Snippet: Effect of EDTA and DNaseI in combination with antibiotics on biofilm cell viability. Biofilms were formed in the flow cell model of NTHi biofilm development and treated with EDTA or DNaseI, alone or in combination with ampicillin or ciprofloxacin. Bacterial viability was assessed with live/dead staining kit and imaged using CLSM. Quantitative analysis of dead cell biomass was performed using COMSTAT of the proridium idodide channel (A). Data presented as mean ± SEM (*** P

Techniques Used: Flow Cytometry, Staining, Confocal Laser Scanning Microscopy

EDTA or DNaseI enhance the efficacy of ampicillin or ciprofloxacin treatment of established biofilms. Biofilms were formed in the flow cell model of NTHi biofilm development and treated with 25 mmol/L EDTA alone or in combination with either 50 μ mol/L of ampicillin or 1.2 μ mol/L ciprofloxacin (upper) or with DNaseI (100 Kunitz U/mL) alone or in combination with either 50 μ mol/L of ampicillin or 1.2 μ mol/L ciprofloxacin (lower). Biofilm biomass was quantitated from CLSM images with COMSTAT. Data presented as mean ± SEM (**** P
Figure Legend Snippet: EDTA or DNaseI enhance the efficacy of ampicillin or ciprofloxacin treatment of established biofilms. Biofilms were formed in the flow cell model of NTHi biofilm development and treated with 25 mmol/L EDTA alone or in combination with either 50 μ mol/L of ampicillin or 1.2 μ mol/L ciprofloxacin (upper) or with DNaseI (100 Kunitz U/mL) alone or in combination with either 50 μ mol/L of ampicillin or 1.2 μ mol/L ciprofloxacin (lower). Biofilm biomass was quantitated from CLSM images with COMSTAT. Data presented as mean ± SEM (**** P

Techniques Used: Flow Cytometry, Confocal Laser Scanning Microscopy

Cation chelators treat established biofilms. To determine the ability of chelators to treat established biofilms, biofilms were formed using static (A–C) or flow (D–F) models of biofilm development, in media supplemented with 2.5 mmol/L Mg 2+ . Biofilms were then treated for 1 h with 25 mmol/L EDTA (B and E) or received no treatment (A and D). Biofilms were stained with SYTO9® and imaged by CLSM (A, B, D, E). Biofilm biomass was quantitated from CLSM images with COMSTAT (C and F). Scale bar 50 μ m. CLSM, confocal laser scanning microscopy, EDTA, ethylenediaminetetra-acetic acid.
Figure Legend Snippet: Cation chelators treat established biofilms. To determine the ability of chelators to treat established biofilms, biofilms were formed using static (A–C) or flow (D–F) models of biofilm development, in media supplemented with 2.5 mmol/L Mg 2+ . Biofilms were then treated for 1 h with 25 mmol/L EDTA (B and E) or received no treatment (A and D). Biofilms were stained with SYTO9® and imaged by CLSM (A, B, D, E). Biofilm biomass was quantitated from CLSM images with COMSTAT (C and F). Scale bar 50 μ m. CLSM, confocal laser scanning microscopy, EDTA, ethylenediaminetetra-acetic acid.

Techniques Used: Flow Cytometry, Staining, Confocal Laser Scanning Microscopy

Mg 2+ enhances NTHi biofilm formation. Biofilms were formed by NTHi 502 in CDM in the absence (A and D) or presence (B and E) of 2.5 mmol/L Mg 2+ using static (A–C) or flow cell (D–F) models of NTHi biofilm develpment. Biofilms were stained with SYTO9® and imaged by CLSM (A, B, D, E). Biofilm biomass was quantitated from CLSM images with COMSTAT (C and F). Scale bar 50 μ m. NTHi, nontypeable Hemophilus influenzae ; CDM, chemically defined medium pH 9; CLSM, confocal laser scanning microscopy.
Figure Legend Snippet: Mg 2+ enhances NTHi biofilm formation. Biofilms were formed by NTHi 502 in CDM in the absence (A and D) or presence (B and E) of 2.5 mmol/L Mg 2+ using static (A–C) or flow cell (D–F) models of NTHi biofilm develpment. Biofilms were stained with SYTO9® and imaged by CLSM (A, B, D, E). Biofilm biomass was quantitated from CLSM images with COMSTAT (C and F). Scale bar 50 μ m. NTHi, nontypeable Hemophilus influenzae ; CDM, chemically defined medium pH 9; CLSM, confocal laser scanning microscopy.

Techniques Used: Flow Cytometry, Staining, Confocal Laser Scanning Microscopy

11) Product Images from "A Filamentous Hemagglutinin-Like Protein of Xanthomonas axonopodis pv. citri, the Phytopathogen Responsible for Citrus Canker, Is Involved in Bacterial Virulence"

Article Title: A Filamentous Hemagglutinin-Like Protein of Xanthomonas axonopodis pv. citri, the Phytopathogen Responsible for Citrus Canker, Is Involved in Bacterial Virulence

Journal: PLoS ONE

doi: 10.1371/journal.pone.0004358

Biofilm formation of X. axonopodis pv. citri wild type and Δ XacFhaB and Δ XacFhaC strains. (A) Representative photograph of the biofilms formed by the gfp -expressing XacWT, Δ XacFhaB and Δ XacFhaC strains grown statically in 12-well PVC plates with SB medium. (B) Representative photographs of confocal laser scanning microscopy of the bacterial strains grown as in A. Magnification 100×. (C) Representative photographs of scanning electron microscopy of the different strains inoculated at a concentration of 10 5 cfu/ml and grown for a period of 20 days in orange leaves. The upper panels shown the cankers formed by XacWT, Δ XacFhaB and Δ XacFhaC at 153× magnification, the lower panels correspond to a higher magnification (10000×). Arrows indicate the different arrays of cells.
Figure Legend Snippet: Biofilm formation of X. axonopodis pv. citri wild type and Δ XacFhaB and Δ XacFhaC strains. (A) Representative photograph of the biofilms formed by the gfp -expressing XacWT, Δ XacFhaB and Δ XacFhaC strains grown statically in 12-well PVC plates with SB medium. (B) Representative photographs of confocal laser scanning microscopy of the bacterial strains grown as in A. Magnification 100×. (C) Representative photographs of scanning electron microscopy of the different strains inoculated at a concentration of 10 5 cfu/ml and grown for a period of 20 days in orange leaves. The upper panels shown the cankers formed by XacWT, Δ XacFhaB and Δ XacFhaC at 153× magnification, the lower panels correspond to a higher magnification (10000×). Arrows indicate the different arrays of cells.

Techniques Used: Expressing, Confocal Laser Scanning Microscopy, Electron Microscopy, Concentration Assay

12) Product Images from "Chemical Genetic Analysis and Functional Characterization of Staphylococcal Wall Teichoic Acid 2-Epimerases Reveals Unconventional Antibiotic Drug Targets"

Article Title: Chemical Genetic Analysis and Functional Characterization of Staphylococcal Wall Teichoic Acid 2-Epimerases Reveals Unconventional Antibiotic Drug Targets

Journal: PLoS Pathogens

doi: 10.1371/journal.ppat.1005585

WTA is required for biofilm formation in methicillin resistant Staphylococci . For total biofilm quantification, biofilms were grown in triplicates for 24 hours in 96-well plates with or without indicated sub-MIC concentrations of WTA inhibitors for MRSA COL (A,C,E) and MRSE CLB26329 (B,D) strains. Genetic complementation of described mutants was performed using plasmid-based copies of wild-type cap5P (p cap5P ), mnaA Sa (p mnaA Sa ), and mnaA Se (p mnaA Se ) as indicated. Biofilms were stained with safranin and dissolved in glacial acetic acid before OD 564 was measured. Bars represent mean OD, error bars represent standard deviation. For Epi fluorescence microscopy, biofilms of MRSA (C) and MRSE (D) were grown as above in black clear bottom plates and stained with Bac Light Green fluorescent stain. Z-stacks were obtained at 60x magnification. Scale bar = 10 μm.
Figure Legend Snippet: WTA is required for biofilm formation in methicillin resistant Staphylococci . For total biofilm quantification, biofilms were grown in triplicates for 24 hours in 96-well plates with or without indicated sub-MIC concentrations of WTA inhibitors for MRSA COL (A,C,E) and MRSE CLB26329 (B,D) strains. Genetic complementation of described mutants was performed using plasmid-based copies of wild-type cap5P (p cap5P ), mnaA Sa (p mnaA Sa ), and mnaA Se (p mnaA Se ) as indicated. Biofilms were stained with safranin and dissolved in glacial acetic acid before OD 564 was measured. Bars represent mean OD, error bars represent standard deviation. For Epi fluorescence microscopy, biofilms of MRSA (C) and MRSE (D) were grown as above in black clear bottom plates and stained with Bac Light Green fluorescent stain. Z-stacks were obtained at 60x magnification. Scale bar = 10 μm.

Techniques Used: Plasmid Preparation, Staining, Standard Deviation, Fluorescence, Microscopy, BAC Assay

13) Product Images from "Anti-Biofilm and Antivirulence Activities of Metabolites from Plectosphaerella cucumerina against Pseudomonas aeruginosa"

Article Title: Anti-Biofilm and Antivirulence Activities of Metabolites from Plectosphaerella cucumerina against Pseudomonas aeruginosa

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2017.00769

Microscopic images of bacterial biofilms grown in the absence and presence of P. cucumerina extract. (A) Light microscopic images and (B) fluorescence microscopic images of (a) distilled water, (b) MeOH, and (c–f) extract (0.25–1 mg mL −1 ) treated biofilms of P. aeruginosa PAO1.
Figure Legend Snippet: Microscopic images of bacterial biofilms grown in the absence and presence of P. cucumerina extract. (A) Light microscopic images and (B) fluorescence microscopic images of (a) distilled water, (b) MeOH, and (c–f) extract (0.25–1 mg mL −1 ) treated biofilms of P. aeruginosa PAO1.

Techniques Used: Fluorescence

Effects of P. cucumerina extract on biofilm development and exopolysaccharide production. (A) Biofilm development and (B) exopolysaccharide production were quantified after 24 h of incubation by measuring at OD 570 and 490 nm, respectively. Error bars indicated the standard deviations of three measurements. Statistically difference was determined by ANOVA followed by Tukey–Kramer test. *** p
Figure Legend Snippet: Effects of P. cucumerina extract on biofilm development and exopolysaccharide production. (A) Biofilm development and (B) exopolysaccharide production were quantified after 24 h of incubation by measuring at OD 570 and 490 nm, respectively. Error bars indicated the standard deviations of three measurements. Statistically difference was determined by ANOVA followed by Tukey–Kramer test. *** p

Techniques Used: Incubation

Effect of P. cucumerina extract on biofilm disruption . (A) Quantitative assessment of biofilm biomass disruption. Mean values of triplicate independent experiments and SD were shown. Statistically difference was determined by ANOVA followed by Tukey–Kramer test. *** p
Figure Legend Snippet: Effect of P. cucumerina extract on biofilm disruption . (A) Quantitative assessment of biofilm biomass disruption. Mean values of triplicate independent experiments and SD were shown. Statistically difference was determined by ANOVA followed by Tukey–Kramer test. *** p

Techniques Used:

Effects of emodin (25 and 50 μg mL −1 ) and patulin (25 and 50 μg mL −1 ) on biofilm development and virulence factors. (A) Biofilm development was quantified after 24 h of incubation by measuring at OD 570. (B) Fluorescence microscopic images of (a) distilled water, (b) MeOH, (c,d) emodin (25 and 50 μg mL −1 , respectively), and (e,f) patulin (25 and 50 μg mL −1 , respectively) treated biofilms of P. aeruginosa PAO1. (C) Quantitative assessment of biofilm biomass disruption. (D) SEM images of (a) distilled water, (b) MeOH, (c,d) emodin (25 and 50 μg mL −1 , respectively), and (e,f) patulin (25 and 50 μg mL −1 , respectively) treated biofilms of P. aeruginosa PAO1. Scale bars = 5 μm. (E) Protease activity. (F) Elastase activity. (G) Effects of emodin on pyocyanin production of P. aeruginosa PAO1. (H) Effects of patulin on pyocyanin production of P. aeruginosa PAO1. (I) Swimming motility assays were performed on plates in the presence of (a) distilled water, (b) MeOH, (c,d) emodin (25 and 50 μg mL −1 , respectively), and (e,f) patulin (25 and 50 μg mL −1 , respectively). Error bars indicated the standard deviations of three measurements. Statistically difference was determined by ANOVA followed by Tukey–Kramer test. ** p
Figure Legend Snippet: Effects of emodin (25 and 50 μg mL −1 ) and patulin (25 and 50 μg mL −1 ) on biofilm development and virulence factors. (A) Biofilm development was quantified after 24 h of incubation by measuring at OD 570. (B) Fluorescence microscopic images of (a) distilled water, (b) MeOH, (c,d) emodin (25 and 50 μg mL −1 , respectively), and (e,f) patulin (25 and 50 μg mL −1 , respectively) treated biofilms of P. aeruginosa PAO1. (C) Quantitative assessment of biofilm biomass disruption. (D) SEM images of (a) distilled water, (b) MeOH, (c,d) emodin (25 and 50 μg mL −1 , respectively), and (e,f) patulin (25 and 50 μg mL −1 , respectively) treated biofilms of P. aeruginosa PAO1. Scale bars = 5 μm. (E) Protease activity. (F) Elastase activity. (G) Effects of emodin on pyocyanin production of P. aeruginosa PAO1. (H) Effects of patulin on pyocyanin production of P. aeruginosa PAO1. (I) Swimming motility assays were performed on plates in the presence of (a) distilled water, (b) MeOH, (c,d) emodin (25 and 50 μg mL −1 , respectively), and (e,f) patulin (25 and 50 μg mL −1 , respectively). Error bars indicated the standard deviations of three measurements. Statistically difference was determined by ANOVA followed by Tukey–Kramer test. ** p

Techniques Used: Incubation, Fluorescence, Activity Assay

14) Product Images from "Manuka-type honeys can eradicate biofilms produced by Staphylococcus aureus strains with different biofilm-forming abilities"

Article Title: Manuka-type honeys can eradicate biofilms produced by Staphylococcus aureus strains with different biofilm-forming abilities

Journal: PeerJ

doi: 10.7717/peerj.326

Effects of NZ honeys and sugar on S. aureus biofilm formation. S. aureus biofilms were allowed to form in the presence of four different NZ honey types (manuka, Medihoney, manuka/kanuka or clover) or a sugar solution. Biofilm formation was assessed using a static biofilm formation assay with crystal violet staining to quantify biomass. S. aureus strains are: (A) NCTC 8325; (B) ATCC 25923; (C) MW2 (HA-MRSA) and (D) USA300 (CA-MRSA). Biofilm formation is expressed as a percentage relative to that produced by the untreated control, which is set at 100%. Error bars represent ±standard deviation (SD) of three biological samples performed in triplicate. Statistical significance ( p
Figure Legend Snippet: Effects of NZ honeys and sugar on S. aureus biofilm formation. S. aureus biofilms were allowed to form in the presence of four different NZ honey types (manuka, Medihoney, manuka/kanuka or clover) or a sugar solution. Biofilm formation was assessed using a static biofilm formation assay with crystal violet staining to quantify biomass. S. aureus strains are: (A) NCTC 8325; (B) ATCC 25923; (C) MW2 (HA-MRSA) and (D) USA300 (CA-MRSA). Biofilm formation is expressed as a percentage relative to that produced by the untreated control, which is set at 100%. Error bars represent ±standard deviation (SD) of three biological samples performed in triplicate. Statistical significance ( p

Techniques Used: Tube Formation Assay, Staining, Produced, Standard Deviation

Correlation of levels of intracellular ATP to colony forming units (CFU) in static biofilms of S. aureus. Static biofilms of S. aureus were formed in the wells of a microtitre plate for 48 h (with media replenishment at 24 h). After removal of the biofilm from the wall of each well, intracellular ATP levels were measured by the BacTitre Glo Viability Kit and CFU were determined for each well. The intracellular levels of ATP are plotted as a function of CFU and validate that the BacTitre Glo Viability Kit can be used as a surrogate measure of biofilm cell viability in subsequent assays.
Figure Legend Snippet: Correlation of levels of intracellular ATP to colony forming units (CFU) in static biofilms of S. aureus. Static biofilms of S. aureus were formed in the wells of a microtitre plate for 48 h (with media replenishment at 24 h). After removal of the biofilm from the wall of each well, intracellular ATP levels were measured by the BacTitre Glo Viability Kit and CFU were determined for each well. The intracellular levels of ATP are plotted as a function of CFU and validate that the BacTitre Glo Viability Kit can be used as a surrogate measure of biofilm cell viability in subsequent assays.

Techniques Used:

Effects of MGO on established S. aureus biofilms. S. aureus NCTC 8325 biofilms were treated with MGO and a combination of MGO and the sugar solution. MGO stock solutions were prepared to correspond to the MGO levels in undiluted honey (100 mg/kg of manuka/kanuka honey, 700 mg/kg of Medihoney, and 900 mg/kg of manuka honey; Table 1 ). The crystal violet stained residual biofilm mass after 24 h treatment was quantified using optical density (OD 595 nm ). Error bars represent ±standard deviation (SD) of three biological samples performed in triplicate. Statistical significance ( p
Figure Legend Snippet: Effects of MGO on established S. aureus biofilms. S. aureus NCTC 8325 biofilms were treated with MGO and a combination of MGO and the sugar solution. MGO stock solutions were prepared to correspond to the MGO levels in undiluted honey (100 mg/kg of manuka/kanuka honey, 700 mg/kg of Medihoney, and 900 mg/kg of manuka honey; Table 1 ). The crystal violet stained residual biofilm mass after 24 h treatment was quantified using optical density (OD 595 nm ). Error bars represent ±standard deviation (SD) of three biological samples performed in triplicate. Statistical significance ( p

Techniques Used: Staining, Standard Deviation

Effects of NZ honeys on established S. aureus biofilms and cell viability within the biofilms. Established S. aureus NCTC 8325, ATCC 25923, MW2 and USA300 biofilms were treated with NZ honeys – manuka, Medihoney, manuka/kanuka, clover, and a sugar solution. The remaining biofilm masses were quantified using crystal violet staining (left y -axis) and cell viability within these remaining biofilms were assessed using the BacTitre Glo Viability Kit (right y -axis). Error bars represent ± standard deviation (SD) of three biological samples performed in triplicate Statistical significance ( p
Figure Legend Snippet: Effects of NZ honeys on established S. aureus biofilms and cell viability within the biofilms. Established S. aureus NCTC 8325, ATCC 25923, MW2 and USA300 biofilms were treated with NZ honeys – manuka, Medihoney, manuka/kanuka, clover, and a sugar solution. The remaining biofilm masses were quantified using crystal violet staining (left y -axis) and cell viability within these remaining biofilms were assessed using the BacTitre Glo Viability Kit (right y -axis). Error bars represent ± standard deviation (SD) of three biological samples performed in triplicate Statistical significance ( p

Techniques Used: Staining, Standard Deviation

Live/dead staining of different honey treated established biofilms. Established biofilms produced by S. aureus NCTC 8325 were treated with TSB containing honey (manuka, Medihoney, manuka/kanuka or clover) or sugar solution at 1%, 2%, 16%, and 32% (w/vol). Syto9 (green; viable cells) and propidium iodine (red; dead cells) stained images were acquired using Nikon A1 Confocal Laser Scanning Microscope. The 3D- images were reconstructed using NIS-elements (version 10). Scale bar represents 50 µ m.
Figure Legend Snippet: Live/dead staining of different honey treated established biofilms. Established biofilms produced by S. aureus NCTC 8325 were treated with TSB containing honey (manuka, Medihoney, manuka/kanuka or clover) or sugar solution at 1%, 2%, 16%, and 32% (w/vol). Syto9 (green; viable cells) and propidium iodine (red; dead cells) stained images were acquired using Nikon A1 Confocal Laser Scanning Microscope. The 3D- images were reconstructed using NIS-elements (version 10). Scale bar represents 50 µ m.

Techniques Used: Staining, Produced, Laser-Scanning Microscopy

Quantitative analysis of live/dead stained honey treated biofilms. The established S. aureus NCTC 8325 biofilm was treated with New Zealand honeys (manuka honey, Medihoney, manuka/kanuka honey, and clover honey) and a sugar solution at 1%, 2%, 16%, and 32% (w/v) concentrations. Biofilms were co-stained with Syto9 (S, viable cells) and propidium iodide (P, dead cells) and analyzed using COMSTAT. The estimated live (S) and dead (P) biomass (volume of the biofilm over the surface area (µm 3 /µm 2 )) are expressed as a percentage of the non-treated control live and dead biomass, which is set at 100%. Error bars represent ±standard deviation (SD) of three biological samples where eight representative images were acquired. Statistical significance ( p
Figure Legend Snippet: Quantitative analysis of live/dead stained honey treated biofilms. The established S. aureus NCTC 8325 biofilm was treated with New Zealand honeys (manuka honey, Medihoney, manuka/kanuka honey, and clover honey) and a sugar solution at 1%, 2%, 16%, and 32% (w/v) concentrations. Biofilms were co-stained with Syto9 (S, viable cells) and propidium iodide (P, dead cells) and analyzed using COMSTAT. The estimated live (S) and dead (P) biomass (volume of the biofilm over the surface area (µm 3 /µm 2 )) are expressed as a percentage of the non-treated control live and dead biomass, which is set at 100%. Error bars represent ±standard deviation (SD) of three biological samples where eight representative images were acquired. Statistical significance ( p

Techniques Used: Staining, Standard Deviation

Quantification of biofilm formation by different strains of S. aureus. The ability of different strains of S. aureus to form biofilms on the plastic surface of tissue-culture treated 96-well microtitre plate was assessed in TSB(G) at 24 h and 48 h. Biofilm adherence was determined using a static biofilm formation assay over 24 h (A) and 48 h (with media replenished after 24 h incubation) (B). Biofilm formation was quantified by staining with 0.2% crystal violet solution and measured at an optical density of 595 nm. Error bars represent ±standard deviation (SD) of three biological samples performed in triplicate, *** represents p
Figure Legend Snippet: Quantification of biofilm formation by different strains of S. aureus. The ability of different strains of S. aureus to form biofilms on the plastic surface of tissue-culture treated 96-well microtitre plate was assessed in TSB(G) at 24 h and 48 h. Biofilm adherence was determined using a static biofilm formation assay over 24 h (A) and 48 h (with media replenished after 24 h incubation) (B). Biofilm formation was quantified by staining with 0.2% crystal violet solution and measured at an optical density of 595 nm. Error bars represent ±standard deviation (SD) of three biological samples performed in triplicate, *** represents p

Techniques Used: Tube Formation Assay, Incubation, Staining, Standard Deviation

Effects of MGO on S. aureus biofilm formation. Biofilm formation by S. aureus NCTC 8325 grown in the presence of MGO and MGO plus sugar solution. MGO stock solutions were prepared to correspond to the MGO levels in undiluted manuka-type honeys (100 mg/kg of manuka/kanuka honey, 700 mg/kg of Medihoney, and 900 mg/kg of manuka-honey; Table 1 ). Biofilm formation was assessed using the described static assay with crystal violet staining to quantify biomass. Biofilm formation is expressed as a percentage relative to the untreated control, which is set at 100%. Error bars represent ±standard deviation (SD) of three biological samples performed in triplicate. Statistical significance ( p
Figure Legend Snippet: Effects of MGO on S. aureus biofilm formation. Biofilm formation by S. aureus NCTC 8325 grown in the presence of MGO and MGO plus sugar solution. MGO stock solutions were prepared to correspond to the MGO levels in undiluted manuka-type honeys (100 mg/kg of manuka/kanuka honey, 700 mg/kg of Medihoney, and 900 mg/kg of manuka-honey; Table 1 ). Biofilm formation was assessed using the described static assay with crystal violet staining to quantify biomass. Biofilm formation is expressed as a percentage relative to the untreated control, which is set at 100%. Error bars represent ±standard deviation (SD) of three biological samples performed in triplicate. Statistical significance ( p

Techniques Used: Staining, Standard Deviation

15) Product Images from "Antibiotic susceptibility of coagulase-negative staphylococci isolated from very low birth weight babies: comprehensive comparisons of bacteria at different stages of biofilm formation"

Article Title: Antibiotic susceptibility of coagulase-negative staphylococci isolated from very low birth weight babies: comprehensive comparisons of bacteria at different stages of biofilm formation

Journal: Annals of Clinical Microbiology and Antimicrobials

doi: 10.1186/1476-0711-9-16

Enhancement of the biofilm growth of isolate 15 by oxacillin at the highest achievable serum concentration* *: As isolate 15 and 22 showed similar responses, only the results for isolate 15 are shown. a and c: stained with Alexa Fluor 555 conjugated wheat germ agglutinin; red signal indicates the presence of polysaccharide. b and d: stained with Live/Dead BacLight bacterial viability kit; green signal indicates the presence of live cells and red signal indicates the presence of dead cells.
Figure Legend Snippet: Enhancement of the biofilm growth of isolate 15 by oxacillin at the highest achievable serum concentration* *: As isolate 15 and 22 showed similar responses, only the results for isolate 15 are shown. a and c: stained with Alexa Fluor 555 conjugated wheat germ agglutinin; red signal indicates the presence of polysaccharide. b and d: stained with Live/Dead BacLight bacterial viability kit; green signal indicates the presence of live cells and red signal indicates the presence of dead cells.

Techniques Used: Concentration Assay, Staining

Enhancement of biofilm-mode growth of CoNS cells by oxacillin at the highest serum achievable concentration. Solutions of gentamicin, oxacillin, vancomycin or the three agents in combination in TSB at the highest concentrations achievable in serum, were added to biofilm grown cells of two S. capitis isolates; 15 (biofilm-negative, ica -weak), and 22 (biofilm-negative, ica -positive). After overnight incubation, bacterial growth was stained with crystal violet and the OD 600 was measured. Error bars represent standard errors of at least 3 individual experiments in triplicate. *: P = 0.004; ** P
Figure Legend Snippet: Enhancement of biofilm-mode growth of CoNS cells by oxacillin at the highest serum achievable concentration. Solutions of gentamicin, oxacillin, vancomycin or the three agents in combination in TSB at the highest concentrations achievable in serum, were added to biofilm grown cells of two S. capitis isolates; 15 (biofilm-negative, ica -weak), and 22 (biofilm-negative, ica -positive). After overnight incubation, bacterial growth was stained with crystal violet and the OD 600 was measured. Error bars represent standard errors of at least 3 individual experiments in triplicate. *: P = 0.004; ** P

Techniques Used: Concentration Assay, Incubation, Staining

16) Product Images from "Anti-Biofilm and Antivirulence Activities of Metabolites from Plectosphaerella cucumerina against Pseudomonas aeruginosa"

Article Title: Anti-Biofilm and Antivirulence Activities of Metabolites from Plectosphaerella cucumerina against Pseudomonas aeruginosa

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2017.00769

Microscopic images of bacterial biofilms grown in the absence and presence of P. cucumerina extract. (A) Light microscopic images and (B) fluorescence microscopic images of (a) distilled water, (b) MeOH, and (c–f) extract (0.25–1 mg mL −1 ) treated biofilms of P. aeruginosa PAO1.
Figure Legend Snippet: Microscopic images of bacterial biofilms grown in the absence and presence of P. cucumerina extract. (A) Light microscopic images and (B) fluorescence microscopic images of (a) distilled water, (b) MeOH, and (c–f) extract (0.25–1 mg mL −1 ) treated biofilms of P. aeruginosa PAO1.

Techniques Used: Fluorescence

Effects of P. cucumerina extract on biofilm development and exopolysaccharide production. (A) Biofilm development and (B) exopolysaccharide production were quantified after 24 h of incubation by measuring at OD 570 and 490 nm, respectively. Error bars indicated the standard deviations of three measurements. Statistically difference was determined by ANOVA followed by Tukey–Kramer test. *** p
Figure Legend Snippet: Effects of P. cucumerina extract on biofilm development and exopolysaccharide production. (A) Biofilm development and (B) exopolysaccharide production were quantified after 24 h of incubation by measuring at OD 570 and 490 nm, respectively. Error bars indicated the standard deviations of three measurements. Statistically difference was determined by ANOVA followed by Tukey–Kramer test. *** p

Techniques Used: Incubation

Effect of P. cucumerina extract on biofilm disruption . (A) Quantitative assessment of biofilm biomass disruption. Mean values of triplicate independent experiments and SD were shown. Statistically difference was determined by ANOVA followed by Tukey–Kramer test. *** p
Figure Legend Snippet: Effect of P. cucumerina extract on biofilm disruption . (A) Quantitative assessment of biofilm biomass disruption. Mean values of triplicate independent experiments and SD were shown. Statistically difference was determined by ANOVA followed by Tukey–Kramer test. *** p

Techniques Used:

Effects of emodin (25 and 50 μg mL −1 ) and patulin (25 and 50 μg mL −1 ) on biofilm development and virulence factors. (A) Biofilm development was quantified after 24 h of incubation by measuring at OD 570. (B) Fluorescence microscopic images of (a) distilled water, (b) MeOH, (c,d) emodin (25 and 50 μg mL −1 , respectively), and (e,f) patulin (25 and 50 μg mL −1 , respectively) treated biofilms of P. aeruginosa PAO1. (C) Quantitative assessment of biofilm biomass disruption. (D) SEM images of (a) distilled water, (b) MeOH, (c,d) emodin (25 and 50 μg mL −1 , respectively), and (e,f) patulin (25 and 50 μg mL −1 , respectively) treated biofilms of P. aeruginosa PAO1. Scale bars = 5 μm. (E) Protease activity. (F) Elastase activity. (G) Effects of emodin on pyocyanin production of P. aeruginosa PAO1. (H) Effects of patulin on pyocyanin production of P. aeruginosa PAO1. (I) Swimming motility assays were performed on plates in the presence of (a) distilled water, (b) MeOH, (c,d) emodin (25 and 50 μg mL −1 , respectively), and (e,f) patulin (25 and 50 μg mL −1 , respectively). Error bars indicated the standard deviations of three measurements. Statistically difference was determined by ANOVA followed by Tukey–Kramer test. ** p
Figure Legend Snippet: Effects of emodin (25 and 50 μg mL −1 ) and patulin (25 and 50 μg mL −1 ) on biofilm development and virulence factors. (A) Biofilm development was quantified after 24 h of incubation by measuring at OD 570. (B) Fluorescence microscopic images of (a) distilled water, (b) MeOH, (c,d) emodin (25 and 50 μg mL −1 , respectively), and (e,f) patulin (25 and 50 μg mL −1 , respectively) treated biofilms of P. aeruginosa PAO1. (C) Quantitative assessment of biofilm biomass disruption. (D) SEM images of (a) distilled water, (b) MeOH, (c,d) emodin (25 and 50 μg mL −1 , respectively), and (e,f) patulin (25 and 50 μg mL −1 , respectively) treated biofilms of P. aeruginosa PAO1. Scale bars = 5 μm. (E) Protease activity. (F) Elastase activity. (G) Effects of emodin on pyocyanin production of P. aeruginosa PAO1. (H) Effects of patulin on pyocyanin production of P. aeruginosa PAO1. (I) Swimming motility assays were performed on plates in the presence of (a) distilled water, (b) MeOH, (c,d) emodin (25 and 50 μg mL −1 , respectively), and (e,f) patulin (25 and 50 μg mL −1 , respectively). Error bars indicated the standard deviations of three measurements. Statistically difference was determined by ANOVA followed by Tukey–Kramer test. ** p

Techniques Used: Incubation, Fluorescence, Activity Assay

17) Product Images from "Baicalin inhibits biofilm formation, attenuates the quorum sensing-controlled virulence and enhances Pseudomonas aeruginosa clearance in a mouse peritoneal implant infection model"

Article Title: Baicalin inhibits biofilm formation, attenuates the quorum sensing-controlled virulence and enhances Pseudomonas aeruginosa clearance in a mouse peritoneal implant infection model

Journal: PLoS ONE

doi: 10.1371/journal.pone.0176883

Inhibitory effects of baicalin on P . aeruginosa PAO1 biofilm formation. In the dose-dependent analysis ( A ), bacterial suspensions were seeded in 96-well flat-bottomed polystyrene microtiter plates exposed to sub-MICs of baicalin (16, 32, 64, 128, and 256 μg/mL) for 24 h, and biofilm mass formation and bacterial counts were quantified in triplicate. Values represent the mean ± standard deviation. * P
Figure Legend Snippet: Inhibitory effects of baicalin on P . aeruginosa PAO1 biofilm formation. In the dose-dependent analysis ( A ), bacterial suspensions were seeded in 96-well flat-bottomed polystyrene microtiter plates exposed to sub-MICs of baicalin (16, 32, 64, 128, and 256 μg/mL) for 24 h, and biofilm mass formation and bacterial counts were quantified in triplicate. Values represent the mean ± standard deviation. * P

Techniques Used: Standard Deviation

Baicalin as a co-treatment in combination with three conventional antibiotics to treat 24-h P . aeruginosa PAO1 biofilms. (A) P . aeruginosa PAO1 biofilm mass and bacterial counts were quantified after treating pre-existing 24-h biofilms with levofloxacin (1 μg/mL), tobramycin (8 μg/mL) and ceftazidime (2 μg/mL) alone or in combination with various sub-MICs (64, 128, and 256 μg/mL) of baicalin for 24 h. (B) Biofilms were exposed to baicalin (256 μg/mL), levofloxacin (1 μg/mL), tobramycin (8 μg/mL), ceftazidime (2 μg/mL) or a baicalin/antibiotic mixture for 6, 12 and 24 h, and changes in biofilm mass formation and bacterial counts were monitored over time. Experiments were performed in triplicate, and values represent the mean ± standard deviation. * P
Figure Legend Snippet: Baicalin as a co-treatment in combination with three conventional antibiotics to treat 24-h P . aeruginosa PAO1 biofilms. (A) P . aeruginosa PAO1 biofilm mass and bacterial counts were quantified after treating pre-existing 24-h biofilms with levofloxacin (1 μg/mL), tobramycin (8 μg/mL) and ceftazidime (2 μg/mL) alone or in combination with various sub-MICs (64, 128, and 256 μg/mL) of baicalin for 24 h. (B) Biofilms were exposed to baicalin (256 μg/mL), levofloxacin (1 μg/mL), tobramycin (8 μg/mL), ceftazidime (2 μg/mL) or a baicalin/antibiotic mixture for 6, 12 and 24 h, and changes in biofilm mass formation and bacterial counts were monitored over time. Experiments were performed in triplicate, and values represent the mean ± standard deviation. * P

Techniques Used: Standard Deviation

Baicalin as a co-treatment in combination with three conventional antibiotics to treat 96-h P . aeruginosa PAO1 biofilms. (A) P . aeruginosa PAO1 biofilm mass and bacterial counts were quantified after treating pre-existing 96-h biofilms with sub-MIC (256 μg/mL) of baicalin for another 24 h. Experiments were performed in triplicate, and values represent the mean ± standard deviation. ** P
Figure Legend Snippet: Baicalin as a co-treatment in combination with three conventional antibiotics to treat 96-h P . aeruginosa PAO1 biofilms. (A) P . aeruginosa PAO1 biofilm mass and bacterial counts were quantified after treating pre-existing 96-h biofilms with sub-MIC (256 μg/mL) of baicalin for another 24 h. Experiments were performed in triplicate, and values represent the mean ± standard deviation. ** P

Techniques Used: Standard Deviation

18) Product Images from "Exploring the Anti-quorum Sensing and Antibiofilm Efficacy of Phytol against Serratia marcescens Associated Acute Pyelonephritis Infection in Wistar Rats"

Article Title: Exploring the Anti-quorum Sensing and Antibiofilm Efficacy of Phytol against Serratia marcescens Associated Acute Pyelonephritis Infection in Wistar Rats

Journal: Frontiers in Cellular and Infection Microbiology

doi: 10.3389/fcimb.2017.00498

Effect of phytol treatment on the virulence gene expression of S. marcescens . Phytol treatment (10 μg/ml) modulated the expression of QS controlled genes involved in virulence factors production and biofilm formation in S. marcescens . Error bar indicates standard deviations from the mean. Student- t test was used to compare the control and treated samples. * indicates significant at p ≤ 0.005, ** Indicates significant at p ≤ 0.001, and *** indicates significant at p ≤ 0.0005.
Figure Legend Snippet: Effect of phytol treatment on the virulence gene expression of S. marcescens . Phytol treatment (10 μg/ml) modulated the expression of QS controlled genes involved in virulence factors production and biofilm formation in S. marcescens . Error bar indicates standard deviations from the mean. Student- t test was used to compare the control and treated samples. * indicates significant at p ≤ 0.005, ** Indicates significant at p ≤ 0.001, and *** indicates significant at p ≤ 0.0005.

Techniques Used: Expressing

Effect of phytol on S. marcescens biofilm formation and growth. Phytol treatment (5 and 10 μg/ml) significantly inhibited the biofilm formation (A) , without affecting the growth of S. marcescens (B) . One percent of methanol was used as vehicle control. Error bar indicates standard deviations from the mean. Student- t test was used to compare the control and treated samples. * Indicates significant at p ≤ 0.005.
Figure Legend Snippet: Effect of phytol on S. marcescens biofilm formation and growth. Phytol treatment (5 and 10 μg/ml) significantly inhibited the biofilm formation (A) , without affecting the growth of S. marcescens (B) . One percent of methanol was used as vehicle control. Error bar indicates standard deviations from the mean. Student- t test was used to compare the control and treated samples. * Indicates significant at p ≤ 0.005.

Techniques Used:

Microscopic analyses of S. marcescens biofilm formation. Light microscopic (A) and CLSM (B) images of phytol treatment slides (5 and 10 μg/ml) showed disintegration of S. marcescens biofilm formation compared to their untreated controls. Effect of phytol on S. marcescens swarming motility: The control plate exhibited extensive swarming motility on soft agar. In contrast, the phytol treatment (5 and 10 μg/ml) considerably inhibited the S. marcescens swarming motility (C) . One percent of methanol was used as a vehicle control.
Figure Legend Snippet: Microscopic analyses of S. marcescens biofilm formation. Light microscopic (A) and CLSM (B) images of phytol treatment slides (5 and 10 μg/ml) showed disintegration of S. marcescens biofilm formation compared to their untreated controls. Effect of phytol on S. marcescens swarming motility: The control plate exhibited extensive swarming motility on soft agar. In contrast, the phytol treatment (5 and 10 μg/ml) considerably inhibited the S. marcescens swarming motility (C) . One percent of methanol was used as a vehicle control.

Techniques Used: Confocal Laser Scanning Microscopy

19) Product Images from "Enhancement of Vancomycin Activity against Biofilms by Using Ultrasound-Targeted Microbubble Destruction ▿"

Article Title: Enhancement of Vancomycin Activity against Biofilms by Using Ultrasound-Targeted Microbubble Destruction ▿

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.00542-11

(A to F) Three-dimensional structural images of biofilms. The biofilms of S. epidermidis RP62A in FluoroDishes were visualized by CLSM with the LIVE/DEAD viability stain (SYTO9/PI). Viable cells exhibit green fluorescence, whereas dead cells exhibit red
Figure Legend Snippet: (A to F) Three-dimensional structural images of biofilms. The biofilms of S. epidermidis RP62A in FluoroDishes were visualized by CLSM with the LIVE/DEAD viability stain (SYTO9/PI). Viable cells exhibit green fluorescence, whereas dead cells exhibit red

Techniques Used: Confocal Laser Scanning Microscopy, Staining, Fluorescence

Comparison of viable bacteria recovered from biofilms and planktonic bacteria after treatments. Data are expressed as means plus standard deviations ( n = 9). (a) Viable counts recovered from biofilms. (b) Viable counts from planktonic cells. The error
Figure Legend Snippet: Comparison of viable bacteria recovered from biofilms and planktonic bacteria after treatments. Data are expressed as means plus standard deviations ( n = 9). (a) Viable counts recovered from biofilms. (b) Viable counts from planktonic cells. The error

Techniques Used:

Schematic diagram of biofilm infection in a rabbit. Biofilm-infected polyethylene dishes were implanted subcutaneously bilateral to the spine, with three dishes on each side.
Figure Legend Snippet: Schematic diagram of biofilm infection in a rabbit. Biofilm-infected polyethylene dishes were implanted subcutaneously bilateral to the spine, with three dishes on each side.

Techniques Used: Infection

Schematic diagram of biofilm treatment. Ultrasound is transmitted though the bottom of the wells via coupling gels.
Figure Legend Snippet: Schematic diagram of biofilm treatment. Ultrasound is transmitted though the bottom of the wells via coupling gels.

Techniques Used:

Morphology of biofilms examined by macroscopy (MACS) and light microscopy (MICS) (original magnification, ×200). Overnight cultures of bacteria were diluted 1:200 and cultured in 96-well plates (200 μl/well) at 37°C for 12 h, and
Figure Legend Snippet: Morphology of biofilms examined by macroscopy (MACS) and light microscopy (MICS) (original magnification, ×200). Overnight cultures of bacteria were diluted 1:200 and cultured in 96-well plates (200 μl/well) at 37°C for 12 h, and

Techniques Used: Magnetic Cell Separation, Light Microscopy, Cell Culture

Semiquantitative analysis of biofilm density. The biofilms of S. epidermidis RP62A were detected by semiquantitative microtiter plate assay. Briefly, overnight cultures of bacteria were diluted 1:200 and cultured in a 96-well plate (200 μl/well)
Figure Legend Snippet: Semiquantitative analysis of biofilm density. The biofilms of S. epidermidis RP62A were detected by semiquantitative microtiter plate assay. Briefly, overnight cultures of bacteria were diluted 1:200 and cultured in a 96-well plate (200 μl/well)

Techniques Used: Cell Culture

The log 10 numbers of CFU/cm 2 ( y axis) of viable Staphylococcus epidermidis recovered from the implanted dishes in the in vivo rabbit model. Biofilm-infected polyethylene dishes were implanted in six groups of New Zealand White female rabbits. Twenty-four
Figure Legend Snippet: The log 10 numbers of CFU/cm 2 ( y axis) of viable Staphylococcus epidermidis recovered from the implanted dishes in the in vivo rabbit model. Biofilm-infected polyethylene dishes were implanted in six groups of New Zealand White female rabbits. Twenty-four

Techniques Used: In Vivo, Infection

20) Product Images from "Functions of Candida albicans cell wall glycosidases Dfg5p and Dcw1p in biofilm formation and HOG MAPK pathway"

Article Title: Functions of Candida albicans cell wall glycosidases Dfg5p and Dcw1p in biofilm formation and HOG MAPK pathway

Journal: PeerJ

doi: 10.7717/peerj.5685

DFG5 and DCW1 mutant biofilms have morphological defects. (A) WT (SC5314), (B) BWP17, (C) DAY185, (D) ES1, (E) ES195 and (F) ES195+Met/Cys. Biofilms of wild type and mutant strains were cultured for 24 h. Microscopic imaging analysis of the biofilms was performed using light microscopy at 200× magnification. Hyphal morphogenesis appeared to be dramatically reduced in the ES1, ES195 and ES195+Met/Cys mutants. ES195+Met/Cys conditional mutant has enlarged cells, prominent nuclei, cell separation defect and pseudohyphal formation.
Figure Legend Snippet: DFG5 and DCW1 mutant biofilms have morphological defects. (A) WT (SC5314), (B) BWP17, (C) DAY185, (D) ES1, (E) ES195 and (F) ES195+Met/Cys. Biofilms of wild type and mutant strains were cultured for 24 h. Microscopic imaging analysis of the biofilms was performed using light microscopy at 200× magnification. Hyphal morphogenesis appeared to be dramatically reduced in the ES1, ES195 and ES195+Met/Cys mutants. ES195+Met/Cys conditional mutant has enlarged cells, prominent nuclei, cell separation defect and pseudohyphal formation.

Techniques Used: Mutagenesis, Cell Culture, Imaging, Light Microscopy

Biofilm formation is affected in Dfg5p and Dcw1p mutants. Biofilm formation was significantly lower for ES1 mutant and ES195+M/C conditional mutant ( p
Figure Legend Snippet: Biofilm formation is affected in Dfg5p and Dcw1p mutants. Biofilm formation was significantly lower for ES1 mutant and ES195+M/C conditional mutant ( p

Techniques Used: Mutagenesis

21) Product Images from "Role of de-N-acetylase PgaB from Aggregatibacter actinomycetemcomitans in exopolysaccharide export in biofilm mode of growth"

Article Title: Role of de-N-acetylase PgaB from Aggregatibacter actinomycetemcomitans in exopolysaccharide export in biofilm mode of growth

Journal: Molecular oral microbiology

doi: 10.1111/omi.12188

The role of the catalytic domain of PgaB in phenotypic variation and biofilm integrity. (a) Deletion of catalytic domain of PgaB (ΔNpgaB) results in a phenotype with attenuated rugged edges characteristic of IDH 781 but lacking the internal star
Figure Legend Snippet: The role of the catalytic domain of PgaB in phenotypic variation and biofilm integrity. (a) Deletion of catalytic domain of PgaB (ΔNpgaB) results in a phenotype with attenuated rugged edges characteristic of IDH 781 but lacking the internal star

Techniques Used:

Confocal scanning laser microscopic image of biofilms. (a) Biofilm growth was imaged at seven different locations and averaged for analysis. The scale bar is at 20 μm; (b) Cell viability for IDH 781 and ΔNpgaB strains at different time
Figure Legend Snippet: Confocal scanning laser microscopic image of biofilms. (a) Biofilm growth was imaged at seven different locations and averaged for analysis. The scale bar is at 20 μm; (b) Cell viability for IDH 781 and ΔNpgaB strains at different time

Techniques Used:

Immunofluorescence evaluation of biofilms for the production of PNAG. PNAG was visualized using the human mAb F598 and a secondary anti-human IgG conjugated to Alexa 488 (left panel). Images of the same field viewed by DAPI to stain DNA is shown in the
Figure Legend Snippet: Immunofluorescence evaluation of biofilms for the production of PNAG. PNAG was visualized using the human mAb F598 and a secondary anti-human IgG conjugated to Alexa 488 (left panel). Images of the same field viewed by DAPI to stain DNA is shown in the

Techniques Used: Immunofluorescence, Staining

22) Product Images from "Gene regulatory network plasticity predates a switch in function of a conserved transcription regulator"

Article Title: Gene regulatory network plasticity predates a switch in function of a conserved transcription regulator

Journal: eLife

doi: 10.7554/eLife.23250

Ndt80 is required for sporulation but is dispensable for biofilm formation, in K. lactis , P. pastoris , and C. lusitaniae . ( A–C ) Light microscope images of genetically matched wild-type and ndt80 deletion strains (Stars indicate diploid cells that have undergone sporulation) and quantification of the percent of cells exhibiting spores, as measured by microscopy (200 cells counted for each strain). ( D–H ) Confocal scanning laser microscopy images of biofilm formation for genetically matched wild-type and ndt80 deletion strains. Top view of biofilm shown above side view for each, with scale bars representing 25 µm. DOI: http://dx.doi.org/10.7554/eLife.23250.014
Figure Legend Snippet: Ndt80 is required for sporulation but is dispensable for biofilm formation, in K. lactis , P. pastoris , and C. lusitaniae . ( A–C ) Light microscope images of genetically matched wild-type and ndt80 deletion strains (Stars indicate diploid cells that have undergone sporulation) and quantification of the percent of cells exhibiting spores, as measured by microscopy (200 cells counted for each strain). ( D–H ) Confocal scanning laser microscopy images of biofilm formation for genetically matched wild-type and ndt80 deletion strains. Top view of biofilm shown above side view for each, with scale bars representing 25 µm. DOI: http://dx.doi.org/10.7554/eLife.23250.014

Techniques Used: Light Microscopy, Microscopy

23) Product Images from "Functions of Candida albicans cell wall glycosidases Dfg5p and Dcw1p in biofilm formation and HOG MAPK pathway"

Article Title: Functions of Candida albicans cell wall glycosidases Dfg5p and Dcw1p in biofilm formation and HOG MAPK pathway

Journal: PeerJ

doi: 10.7717/peerj.5685

DFG5 and DCW1 mutant biofilms have morphological defects. (A) WT (SC5314), (B) BWP17, (C) DAY185, (D) ES1, (E) ES195 and (F) ES195+Met/Cys. Biofilms of wild type and mutant strains were cultured for 24 h. Microscopic imaging analysis of the biofilms was performed using light microscopy at 200× magnification. Hyphal morphogenesis appeared to be dramatically reduced in the ES1, ES195 and ES195+Met/Cys mutants. ES195+Met/Cys conditional mutant has enlarged cells, prominent nuclei, cell separation defect and pseudohyphal formation.
Figure Legend Snippet: DFG5 and DCW1 mutant biofilms have morphological defects. (A) WT (SC5314), (B) BWP17, (C) DAY185, (D) ES1, (E) ES195 and (F) ES195+Met/Cys. Biofilms of wild type and mutant strains were cultured for 24 h. Microscopic imaging analysis of the biofilms was performed using light microscopy at 200× magnification. Hyphal morphogenesis appeared to be dramatically reduced in the ES1, ES195 and ES195+Met/Cys mutants. ES195+Met/Cys conditional mutant has enlarged cells, prominent nuclei, cell separation defect and pseudohyphal formation.

Techniques Used: Mutagenesis, Cell Culture, Imaging, Light Microscopy

Biofilm formation is affected in Dfg5p and Dcw1p mutants. Biofilm formation was significantly lower for ES1 mutant and ES195+M/C conditional mutant ( p
Figure Legend Snippet: Biofilm formation is affected in Dfg5p and Dcw1p mutants. Biofilm formation was significantly lower for ES1 mutant and ES195+M/C conditional mutant ( p

Techniques Used: Mutagenesis

24) Product Images from "Magnesium Ion Acts as a Signal for Capsule Induction in Cryptococcus neoformans"

Article Title: Magnesium Ion Acts as a Signal for Capsule Induction in Cryptococcus neoformans

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2016.00325

Biofilm formation on coverslip with different media. Biofilm observed at 40× light microscope by India ink stain. Images of a mature biofilm show yeast cells and capsular polysaccharide.
Figure Legend Snippet: Biofilm formation on coverslip with different media. Biofilm observed at 40× light microscope by India ink stain. Images of a mature biofilm show yeast cells and capsular polysaccharide.

Techniques Used: Light Microscopy, Staining

25) Product Images from "Komodo dragon-inspired synthetic peptide DRGN-1 promotes wound-healing of a mixed-biofilm infected wound"

Article Title: Komodo dragon-inspired synthetic peptide DRGN-1 promotes wound-healing of a mixed-biofilm infected wound

Journal: NPJ Biofilms and Microbiomes

doi: 10.1038/s41522-017-0017-2

Efficacy in wound healing of peptides in a mixed biofilm-infected mouse skin wound model. a Schematic experimental design that includes the timing of infection, drug treatments, dressing off. b Effect of peptides on closure of P. aeruginosa/ S. aureus -infected wounds in mice. Wounds shown are representative of the group. Six-millimeter-diameter excisions were made on the back of mice. Each wound was infected with a mixed biofilm ( P. aeruginosa/ S. aureus ) grown on a polycarbonate membrane for 2 days. DRGN-1, VK25 or LL-37 (20 µg) was topically treated every 2 days in 1% hypromellose ( n = 6 mice per group). Mice were lightly anesthetized with isofluorane and photographed on the indicated days following treatment. c , d Assessment of bacterial colonization in P. aeruginosa / S. aureus -infected wounds. Wound tissue samples were harvested on day 6 after treatment, and the number of CFU/wound was counted in a selective medium for each species. The median value in each group is shown as a horizontal bar ( n = 6; ** p
Figure Legend Snippet: Efficacy in wound healing of peptides in a mixed biofilm-infected mouse skin wound model. a Schematic experimental design that includes the timing of infection, drug treatments, dressing off. b Effect of peptides on closure of P. aeruginosa/ S. aureus -infected wounds in mice. Wounds shown are representative of the group. Six-millimeter-diameter excisions were made on the back of mice. Each wound was infected with a mixed biofilm ( P. aeruginosa/ S. aureus ) grown on a polycarbonate membrane for 2 days. DRGN-1, VK25 or LL-37 (20 µg) was topically treated every 2 days in 1% hypromellose ( n = 6 mice per group). Mice were lightly anesthetized with isofluorane and photographed on the indicated days following treatment. c , d Assessment of bacterial colonization in P. aeruginosa / S. aureus -infected wounds. Wound tissue samples were harvested on day 6 after treatment, and the number of CFU/wound was counted in a selective medium for each species. The median value in each group is shown as a horizontal bar ( n = 6; ** p

Techniques Used: Infection, Mouse Assay

Anti-biofilm activity of DRGN-1. a DRGN-1 anti-biofilm activity against S. aureus using three interdependent methods. The activity was analyzed by crystal violet staining of biofilm in culture tube at 24 h. b DRGN-1 anti-biofilm activity against P. aeruginosa . Pictures ( right inlet in a and b ) are a representative biofilm stained with crystal violet. P values were calculated using a two-tailed t test (assuming unequal variances) comparing test strains to untreated control (* P
Figure Legend Snippet: Anti-biofilm activity of DRGN-1. a DRGN-1 anti-biofilm activity against S. aureus using three interdependent methods. The activity was analyzed by crystal violet staining of biofilm in culture tube at 24 h. b DRGN-1 anti-biofilm activity against P. aeruginosa . Pictures ( right inlet in a and b ) are a representative biofilm stained with crystal violet. P values were calculated using a two-tailed t test (assuming unequal variances) comparing test strains to untreated control (* P

Techniques Used: Activity Assay, Staining, Two Tailed Test

26) Product Images from "Absence of TolC Impairs Biofilm Formation in Actinobacillus pleuropneumoniae by Reducing Initial Attachment"

Article Title: Absence of TolC Impairs Biofilm Formation in Actinobacillus pleuropneumoniae by Reducing Initial Attachment

Journal: PLoS ONE

doi: 10.1371/journal.pone.0163364

CLSM images of biofilms formed by WT and Δ tolC . Both 4-h and 6-h static biofilms cultured in 6-well microtiter plates were stained with the SYTO-9 and propidium iodide (A-C) to label live versus dead cells, or with WGA-conjugated to Oregon green to label PGA (D).
Figure Legend Snippet: CLSM images of biofilms formed by WT and Δ tolC . Both 4-h and 6-h static biofilms cultured in 6-well microtiter plates were stained with the SYTO-9 and propidium iodide (A-C) to label live versus dead cells, or with WGA-conjugated to Oregon green to label PGA (D).

Techniques Used: Confocal Laser Scanning Microscopy, Cell Culture, Staining, Whole Genome Amplification

Detachment of biofilm colonies of WT A . pleuropneumoniae and Δ tolC grown in 96-well microtiter plates. Biofilms were cultured for 4h (A), 6h (B) and 12h (C) and treated with dispersin B, DNase I and proteinase K, respectively. The remaining biofilms were quantitated by crystal violet staining. Data represent mean values ± SDs of three independent experiments with three replicates. Results with significant differences when compared to the untreated controls or between two strains were marked with asterisks (P
Figure Legend Snippet: Detachment of biofilm colonies of WT A . pleuropneumoniae and Δ tolC grown in 96-well microtiter plates. Biofilms were cultured for 4h (A), 6h (B) and 12h (C) and treated with dispersin B, DNase I and proteinase K, respectively. The remaining biofilms were quantitated by crystal violet staining. Data represent mean values ± SDs of three independent experiments with three replicates. Results with significant differences when compared to the untreated controls or between two strains were marked with asterisks (P

Techniques Used: Cell Culture, Staining

Effects of tolC deletion on biofilm formation of A . pleuropneumoniae . Biofilm formation was determined at various time points (0, 4, 6, 12, 24 and 36h) by crystal violet staining using static broth cultures of WT SC1516, Δ tolC and the genetically-complemented strain Δ tolC / tolC . Data are means ± standard deviations (SDs) of three independent experiments with three replicates. Asterisks indicate statistical significance (P
Figure Legend Snippet: Effects of tolC deletion on biofilm formation of A . pleuropneumoniae . Biofilm formation was determined at various time points (0, 4, 6, 12, 24 and 36h) by crystal violet staining using static broth cultures of WT SC1516, Δ tolC and the genetically-complemented strain Δ tolC / tolC . Data are means ± standard deviations (SDs) of three independent experiments with three replicates. Asterisks indicate statistical significance (P

Techniques Used: Staining

Coculturing with WT rescued biofilm formation defects of Δ tolC . Biofilm formation was performed by coculturing Δ tolC with WT in Transwell chambers. (A) Biofilms in the lower chamber were stained with crystal violet. (B) Quantitative analysis of biofilms. Data presented here are means of triplicate experiments and error bars indicate the SDs. *, statistical significance (P
Figure Legend Snippet: Coculturing with WT rescued biofilm formation defects of Δ tolC . Biofilm formation was performed by coculturing Δ tolC with WT in Transwell chambers. (A) Biofilms in the lower chamber were stained with crystal violet. (B) Quantitative analysis of biofilms. Data presented here are means of triplicate experiments and error bars indicate the SDs. *, statistical significance (P

Techniques Used: Staining

Relative expression of pgaA and cpxR in biofilms of Δ tolC versus WT. The mRNA levels of pgaA and cpxR genes in biofilms at 4h (A), 6h (B) and 12h (C) were determined by qRT-PCR. Data presented are means of three independent experiments with triplicates. Error bars indicate SDs. Results with statistical significant are marked with asterisks (P
Figure Legend Snippet: Relative expression of pgaA and cpxR in biofilms of Δ tolC versus WT. The mRNA levels of pgaA and cpxR genes in biofilms at 4h (A), 6h (B) and 12h (C) were determined by qRT-PCR. Data presented are means of three independent experiments with triplicates. Error bars indicate SDs. Results with statistical significant are marked with asterisks (P

Techniques Used: Expressing, Quantitative RT-PCR

27) Product Images from "Purification and Characterization of Biofilm-Associated EPS Exopolysaccharides from ESKAPE Organisms and Other Pathogens"

Article Title: Purification and Characterization of Biofilm-Associated EPS Exopolysaccharides from ESKAPE Organisms and Other Pathogens

Journal: PLoS ONE

doi: 10.1371/journal.pone.0067950

EPS staining of E. coli strain 43894 biofilms. The FITC-labeled mannose-specific HHA lectin was used to stain exopolysaccharides (green) and Hoechst 33342 was used to stain the bacterial nucleic acids (blue). ( A. ) Extracellular green staining of the EPS by FITC-HHA can be seen on 1 day old biofilms of E. coli at 200X. Scale bar = 5 µm. ( B. ) Confocal image of 3 day old E. coli biofilms at 63X. The large square panel is a plan view looking down on the biofilm. The top and right-side rectangular panels are vertical sections representing the XZ plane and YZ plane, respectively, at the positions indicated by the colored lines. The biofilm is 40 µm thick (i.e Z-axis).
Figure Legend Snippet: EPS staining of E. coli strain 43894 biofilms. The FITC-labeled mannose-specific HHA lectin was used to stain exopolysaccharides (green) and Hoechst 33342 was used to stain the bacterial nucleic acids (blue). ( A. ) Extracellular green staining of the EPS by FITC-HHA can be seen on 1 day old biofilms of E. coli at 200X. Scale bar = 5 µm. ( B. ) Confocal image of 3 day old E. coli biofilms at 63X. The large square panel is a plan view looking down on the biofilm. The top and right-side rectangular panels are vertical sections representing the XZ plane and YZ plane, respectively, at the positions indicated by the colored lines. The biofilm is 40 µm thick (i.e Z-axis).

Techniques Used: Staining, Labeling

28) Product Images from "A Cardiolipin-Deficient Mutant of Rhodobacter sphaeroides Has an Altered Cell Shape and Is Impaired in Biofilm Formation"

Article Title: A Cardiolipin-Deficient Mutant of Rhodobacter sphaeroides Has an Altered Cell Shape and Is Impaired in Biofilm Formation

Journal: Journal of Bacteriology

doi: 10.1128/JB.00420-15

Complementation of the CL3 strain restores biofilm formation. (A) Images depicting the morphology of R. sphaeroides wild-type (WT) cells harboring the empty vector pIND5sp (Ctrl/WT) and CL3 cells expressing CL synthase from cls -pIND5sp (Cls/CL3). Each
Figure Legend Snippet: Complementation of the CL3 strain restores biofilm formation. (A) Images depicting the morphology of R. sphaeroides wild-type (WT) cells harboring the empty vector pIND5sp (Ctrl/WT) and CL3 cells expressing CL synthase from cls -pIND5sp (Cls/CL3). Each

Techniques Used: Plasmid Preparation, Expressing

R. sphaeroides strain CL3 forms defective biofilms. (A) Representative fluorescence micrographs of R. sphaeroides wild-type (WT) and CL3 biofilms grown on a chamber slide with hydrophobic plastic surfaces for 72 h at 30°C in Sistrom's succinate
Figure Legend Snippet: R. sphaeroides strain CL3 forms defective biofilms. (A) Representative fluorescence micrographs of R. sphaeroides wild-type (WT) and CL3 biofilms grown on a chamber slide with hydrophobic plastic surfaces for 72 h at 30°C in Sistrom's succinate

Techniques Used: Fluorescence

A22 treatment impairs biofilm formation. (A) Images depicting the morphology of R. sphaeroides wild-type (WT) cells treated with DMSO or 10 μg/ml A22. Each datum point was determined by imaging 300 cells by phase-contrast bright-field microscopy
Figure Legend Snippet: A22 treatment impairs biofilm formation. (A) Images depicting the morphology of R. sphaeroides wild-type (WT) cells treated with DMSO or 10 μg/ml A22. Each datum point was determined by imaging 300 cells by phase-contrast bright-field microscopy

Techniques Used: Imaging, Microscopy

29) Product Images from "The LuxS/AI-2 Quorum-Sensing System of Streptococcus pneumoniae Is Required to Cause Disease, and to Regulate Virulence- and Metabolism-Related Genes in a Rat Model of Middle Ear Infection"

Article Title: The LuxS/AI-2 Quorum-Sensing System of Streptococcus pneumoniae Is Required to Cause Disease, and to Regulate Virulence- and Metabolism-Related Genes in a Rat Model of Middle Ear Infection

Journal: Frontiers in Cellular and Infection Microbiology

doi: 10.3389/fcimb.2018.00138

In vitro biofilm growth of Streptococcus pneumoniae D39 wild-type and D39Δ luxS strains at different time-points after inoculation (6, 12, 18, and 24 h). The error bars are the standard deviation from the mean. Statistical significance was calculated using the Student's t -test, * p
Figure Legend Snippet: In vitro biofilm growth of Streptococcus pneumoniae D39 wild-type and D39Δ luxS strains at different time-points after inoculation (6, 12, 18, and 24 h). The error bars are the standard deviation from the mean. Statistical significance was calculated using the Student's t -test, * p

Techniques Used: In Vitro, Standard Deviation

Scanning electron microscopy (SEM) images of Streptococcus pneumoniae in vitro biofilms grown for 18 h. (A–C) are representative SEM images of the D39 wild-type strain. The wild-type strain biofilms were thick and organized with a significant depth. (D–F) are SEM images of the D39Δ luxS strain. The D39Δ luxS biofilms were thin and disorganized, and extracellular polymeric substance (EPS) was absent.
Figure Legend Snippet: Scanning electron microscopy (SEM) images of Streptococcus pneumoniae in vitro biofilms grown for 18 h. (A–C) are representative SEM images of the D39 wild-type strain. The wild-type strain biofilms were thick and organized with a significant depth. (D–F) are SEM images of the D39Δ luxS strain. The D39Δ luxS biofilms were thin and disorganized, and extracellular polymeric substance (EPS) was absent.

Techniques Used: Electron Microscopy, In Vitro

Confocal microscopy images of Streptococcus pneumoniae in vitro biofilms grown for 18 h. (A) Confocal microscopy image of the D39 wild-type strain biofilm. (B) Confocal microscopy image of the D39Δ luxS strain biofilm.
Figure Legend Snippet: Confocal microscopy images of Streptococcus pneumoniae in vitro biofilms grown for 18 h. (A) Confocal microscopy image of the D39 wild-type strain biofilm. (B) Confocal microscopy image of the D39Δ luxS strain biofilm.

Techniques Used: Confocal Microscopy, In Vitro

Streptococcus pneumoniae D39 wild-type and D39Δ luxS planktonic and biofilm growth. (A) Planktonic growth optical density at 600 nm. (B) Quantification of biomass of in vitro biofilms grown for 18 h, using a CV-microplate assay. (C) Colony-forming unit (CFU) counts of in vitro biofilms grown for 18 h. Error bars are the standard deviation from the mean. Statistical significance was calculated using the Student's t -test, * p
Figure Legend Snippet: Streptococcus pneumoniae D39 wild-type and D39Δ luxS planktonic and biofilm growth. (A) Planktonic growth optical density at 600 nm. (B) Quantification of biomass of in vitro biofilms grown for 18 h, using a CV-microplate assay. (C) Colony-forming unit (CFU) counts of in vitro biofilms grown for 18 h. Error bars are the standard deviation from the mean. Statistical significance was calculated using the Student's t -test, * p

Techniques Used: In Vitro, Standard Deviation

30) Product Images from "In silico identification of albendazole as a quorum sensing inhibitor and its in vitro verification using CviR and LasB receptors based assay systems"

Article Title: In silico identification of albendazole as a quorum sensing inhibitor and its in vitro verification using CviR and LasB receptors based assay systems

Journal: BioImpacts : BI

doi: 10.15171/bi.2018.23

Effect of albendazole on Pseudomonas aeruginosa biofilms
Figure Legend Snippet: Effect of albendazole on Pseudomonas aeruginosa biofilms

Techniques Used:

31) Product Images from "3,5-Dicaffeoylquinic Acid Disperses Aspergillus Fumigatus Biofilm and Enhances Fungicidal Efficacy of Voriconazole and Amphotericin B"

Article Title: 3,5-Dicaffeoylquinic Acid Disperses Aspergillus Fumigatus Biofilm and Enhances Fungicidal Efficacy of Voriconazole and Amphotericin B

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

doi: 10.12659/MSM.908068

Metabolic activity alteration of cells in A. fumigatus GXMU04 ( A ) and A. fumigatus AF293 ( B ) of preformed biofilms after exposing to the tested agents evaluated by XTT reduction assay. Error bars represent standard error. * P
Figure Legend Snippet: Metabolic activity alteration of cells in A. fumigatus GXMU04 ( A ) and A. fumigatus AF293 ( B ) of preformed biofilms after exposing to the tested agents evaluated by XTT reduction assay. Error bars represent standard error. * P

Techniques Used: Activity Assay

SEM ( A ) images and CLSM ( B ) view of preformed A. fumigatus GXMU04 biofilms stained with the combination of PI and Syto-9 in control group, MFC of VRC or AMB, 3,5-DCQA (1024 mg/L), and 3,5-DCQA in combination with VRC or AMB.
Figure Legend Snippet: SEM ( A ) images and CLSM ( B ) view of preformed A. fumigatus GXMU04 biofilms stained with the combination of PI and Syto-9 in control group, MFC of VRC or AMB, 3,5-DCQA (1024 mg/L), and 3,5-DCQA in combination with VRC or AMB.

Techniques Used: Confocal Laser Scanning Microscopy, Staining

32) Product Images from "In silico identification of albendazole as a quorum sensing inhibitor and its in vitro verification using CviR and LasB receptors based assay systems"

Article Title: In silico identification of albendazole as a quorum sensing inhibitor and its in vitro verification using CviR and LasB receptors based assay systems

Journal: BioImpacts : BI

doi: 10.15171/bi.2018.23

Effect of albendazole on Pseudomonas aeruginosa biofilms
Figure Legend Snippet: Effect of albendazole on Pseudomonas aeruginosa biofilms

Techniques Used:

33) Product Images from "Polymorphonuclear Leukocytes or Hydrogen Peroxide Enhance Biofilm Development of Mucoid Pseudomonas aeruginosa"

Article Title: Polymorphonuclear Leukocytes or Hydrogen Peroxide Enhance Biofilm Development of Mucoid Pseudomonas aeruginosa

Journal: Mediators of Inflammation

doi: 10.1155/2018/8151362

Effect of PMNs and H 2 O 2 on viable cells of biofilms of mucoid P. aeruginosa FRD1. The error bars indicate standard deviations. Control: control group; PMNs: FRD1 biofilm group treated with PMNs; 1 mM H 2 O 2 : FRD1 biofilm group treated with 1 mM H 2 O 2 ; 2 mM H 2 O 2 : FRD1 biofilm group treated with 2 mM H 2 O 2 . (a, b) The numbers of viable cells in biofilms treated with PMNs are expressed in colony-forming units (CFUs). (c, d) The numbers of viable cells in biofilms treated with H 2 O 2 are expressed in colony-forming units (CFUs). Data are presented as the mean ± SD ( n = 6 in each treatment). ∗ P
Figure Legend Snippet: Effect of PMNs and H 2 O 2 on viable cells of biofilms of mucoid P. aeruginosa FRD1. The error bars indicate standard deviations. Control: control group; PMNs: FRD1 biofilm group treated with PMNs; 1 mM H 2 O 2 : FRD1 biofilm group treated with 1 mM H 2 O 2 ; 2 mM H 2 O 2 : FRD1 biofilm group treated with 2 mM H 2 O 2 . (a, b) The numbers of viable cells in biofilms treated with PMNs are expressed in colony-forming units (CFUs). (c, d) The numbers of viable cells in biofilms treated with H 2 O 2 are expressed in colony-forming units (CFUs). Data are presented as the mean ± SD ( n = 6 in each treatment). ∗ P

Techniques Used:

Effect of PMNs and H 2 O 2 on thickness of biofilms of mucoid P. aeruginosa FRD1. The error bars indicate standard deviations. Control: control group; PMNs: FRD1 biofilm group treated with PMNs; 1 mM H 2 O 2 : FRD1 biofilm group treated with 1 mM H 2 O 2 ; 2 mM H 2 O 2 : FRD1 biofilm group treated with 2 mM H 2 O 2 . (a, b) The thickness of biofilms treated with PMNs was determined using confocal laser-scanning microscopy. (c, d) The thickness of biofilms treated with H 2 O 2 was determined using confocal laser-scanning microscopy. Data are presented as the mean ± SD ( n = 6 in each treatment). ∗ P
Figure Legend Snippet: Effect of PMNs and H 2 O 2 on thickness of biofilms of mucoid P. aeruginosa FRD1. The error bars indicate standard deviations. Control: control group; PMNs: FRD1 biofilm group treated with PMNs; 1 mM H 2 O 2 : FRD1 biofilm group treated with 1 mM H 2 O 2 ; 2 mM H 2 O 2 : FRD1 biofilm group treated with 2 mM H 2 O 2 . (a, b) The thickness of biofilms treated with PMNs was determined using confocal laser-scanning microscopy. (c, d) The thickness of biofilms treated with H 2 O 2 was determined using confocal laser-scanning microscopy. Data are presented as the mean ± SD ( n = 6 in each treatment). ∗ P

Techniques Used: Confocal Laser Scanning Microscopy

Effect of nonactivated PMNs on viable cells of biofilms of mucoid P. aeruginosa FRD1. The numbers of viable cells in biofilms treated with PMNs are expressed in colony-forming units (CFUs). The error bars indicate standard deviations. BF: biofilm without PMNs; BF + nonactivated PMNs: biofilm treated with nonactivated PMNs. Data are presented as the mean ± SD ( n = 6 in each treatment).
Figure Legend Snippet: Effect of nonactivated PMNs on viable cells of biofilms of mucoid P. aeruginosa FRD1. The numbers of viable cells in biofilms treated with PMNs are expressed in colony-forming units (CFUs). The error bars indicate standard deviations. BF: biofilm without PMNs; BF + nonactivated PMNs: biofilm treated with nonactivated PMNs. Data are presented as the mean ± SD ( n = 6 in each treatment).

Techniques Used:

Effect of PMNs on G6PDH activity. G6PDH: glucose-6-phosphate dehydrogenase; Control: P. aeruginosa FRD1 biofilm grown for 24 hours or 48 hours without PMNs; PMN: P. aeruginosa FRD1 biofilm treated with PMNs for 24 hours or 48 hours. The activity of G6PDH did not differ significantly between the two groups. Data are presented as the mean ± SD ( n = 6 in each treatment).
Figure Legend Snippet: Effect of PMNs on G6PDH activity. G6PDH: glucose-6-phosphate dehydrogenase; Control: P. aeruginosa FRD1 biofilm grown for 24 hours or 48 hours without PMNs; PMN: P. aeruginosa FRD1 biofilm treated with PMNs for 24 hours or 48 hours. The activity of G6PDH did not differ significantly between the two groups. Data are presented as the mean ± SD ( n = 6 in each treatment).

Techniques Used: Activity Assay

Effect of PMNs on GMD activity. GMD: GDP-mannose dehydrogenase; Control: P. aeruginosa FRD1 biofilm grown for 24 hours or 48 hours without PMNs; PMN: P. aeruginosa FRD1 biofilm treated with PMNs for 24 hours or 48 hours. Activities of GMD in FRD1 biofilms treated with PMNs were significantly different from those in FRD1 biofilms without PMNs. Data are presented as the means ± SD ( n = 6 in each treatment). ∗ P
Figure Legend Snippet: Effect of PMNs on GMD activity. GMD: GDP-mannose dehydrogenase; Control: P. aeruginosa FRD1 biofilm grown for 24 hours or 48 hours without PMNs; PMN: P. aeruginosa FRD1 biofilm treated with PMNs for 24 hours or 48 hours. Activities of GMD in FRD1 biofilms treated with PMNs were significantly different from those in FRD1 biofilms without PMNs. Data are presented as the means ± SD ( n = 6 in each treatment). ∗ P

Techniques Used: Activity Assay

Effect of PMNs and H 2 O 2 on alginate content. The error bars indicate standard deviations. Control: control group; PMNs: FRD1 biofilm group treated with PMNs; 1 mM H 2 O 2 : FRD1 biofilm group treated with 1 mM H 2 O 2 ; 2 mM H 2 O 2 : FRD1 biofilm group treated with 2 mM H 2 O 2 . (a, b)The alginate content of P. aeruginosa FRD1 biofilms treated with PMNs. (c, d) The alginate content of P. aeruginosa FRD1 biofilms treated with H 2 O 2 . Data are presented as the mean ± SD ( n = 6 in each treatment). ∗ P
Figure Legend Snippet: Effect of PMNs and H 2 O 2 on alginate content. The error bars indicate standard deviations. Control: control group; PMNs: FRD1 biofilm group treated with PMNs; 1 mM H 2 O 2 : FRD1 biofilm group treated with 1 mM H 2 O 2 ; 2 mM H 2 O 2 : FRD1 biofilm group treated with 2 mM H 2 O 2 . (a, b)The alginate content of P. aeruginosa FRD1 biofilms treated with PMNs. (c, d) The alginate content of P. aeruginosa FRD1 biofilms treated with H 2 O 2 . Data are presented as the mean ± SD ( n = 6 in each treatment). ∗ P

Techniques Used:

Effects of PMNs and H 2 O 2 on the expression of genes involved in alginate biosynthesis. Control: control group; PMNs: FRD1 biofilm group treated with PMNs; 1 mM H 2 O 2 : FRD1 biofilm group treated with 1 mM H 2 O 2 ; 2 mM H 2 O 2 : FRD1 biofilm group treated with 2 mM H 2 O 2 . Bacteria were grown in Jensen's medium. RNA was extracted from the bacteria, and the relative mRNA levels of algD , algR , and algU were determined by RT-PCR. (a, b) The expression of genes involved in alginate biosynthesis after treatment with PMNs. (c, d) The expression of genes involved in alginate biosynthesis after treatment with H 2 O 2 . Data are presented as the mean ± SD ( n = 6 in each treatment). ∗ P
Figure Legend Snippet: Effects of PMNs and H 2 O 2 on the expression of genes involved in alginate biosynthesis. Control: control group; PMNs: FRD1 biofilm group treated with PMNs; 1 mM H 2 O 2 : FRD1 biofilm group treated with 1 mM H 2 O 2 ; 2 mM H 2 O 2 : FRD1 biofilm group treated with 2 mM H 2 O 2 . Bacteria were grown in Jensen's medium. RNA was extracted from the bacteria, and the relative mRNA levels of algD , algR , and algU were determined by RT-PCR. (a, b) The expression of genes involved in alginate biosynthesis after treatment with PMNs. (c, d) The expression of genes involved in alginate biosynthesis after treatment with H 2 O 2 . Data are presented as the mean ± SD ( n = 6 in each treatment). ∗ P

Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

Effect of PMA on viable cells of biofilms of mucoid P. aeruginosa FRD1. The numbers of viable cells in biofilms treated with PMA are expressed in colony-forming units (CFUs). The error bars indicate standard deviations. BF: biofilm without PMA; BF + PMA: biofilm treated with PMA (100 ng/ml). Data are presented as the mean ± SD ( n = 6 in each treatment).
Figure Legend Snippet: Effect of PMA on viable cells of biofilms of mucoid P. aeruginosa FRD1. The numbers of viable cells in biofilms treated with PMA are expressed in colony-forming units (CFUs). The error bars indicate standard deviations. BF: biofilm without PMA; BF + PMA: biofilm treated with PMA (100 ng/ml). Data are presented as the mean ± SD ( n = 6 in each treatment).

Techniques Used:

Effect of PMNs and H 2 O 2 on the adhesion of mucoid P. aeruginosa FRD1. The error bars indicate standard deviations. Control: control group; PMNs: FRD1 group treated with PMNs; 1 mM H 2 O 2 : FRD1 group treated with 1 mM H 2 O 2 ; 2 mM H 2 O 2 : FRD1 group treated with 2 mM H 2 O 2 . (a) Quantification of biofilm formation using crystal violet staining. Cells were grown in Jensen's medium for one day, in the presence of PMNs. PMNs increased the adhesion of P. aeruginosa FRD1. (b) The effect of PMNs on the number of viable cells is expressed in colony-forming units (CFUs). (c) Quantification of biofilm formation by crystal violet staining. H 2 O 2 increased the adhesion of P. aeruginosa FRD1. (d) The effect of H 2 O 2 on the number of viable cells is expressed in colony-forming units (CFUs). Data are presented as the mean ± SD ( n = 6 in each treatment). ∗ P
Figure Legend Snippet: Effect of PMNs and H 2 O 2 on the adhesion of mucoid P. aeruginosa FRD1. The error bars indicate standard deviations. Control: control group; PMNs: FRD1 group treated with PMNs; 1 mM H 2 O 2 : FRD1 group treated with 1 mM H 2 O 2 ; 2 mM H 2 O 2 : FRD1 group treated with 2 mM H 2 O 2 . (a) Quantification of biofilm formation using crystal violet staining. Cells were grown in Jensen's medium for one day, in the presence of PMNs. PMNs increased the adhesion of P. aeruginosa FRD1. (b) The effect of PMNs on the number of viable cells is expressed in colony-forming units (CFUs). (c) Quantification of biofilm formation by crystal violet staining. H 2 O 2 increased the adhesion of P. aeruginosa FRD1. (d) The effect of H 2 O 2 on the number of viable cells is expressed in colony-forming units (CFUs). Data are presented as the mean ± SD ( n = 6 in each treatment). ∗ P

Techniques Used: Staining

Effect of PMNs on biofilms of mucoid P. aeruginosa FRD1. Confocal laser scanning micrographs of P. aeruginosa FRD1 biofilms treated with PMNs. (a) Control group, early biofilm treated without PMNs. (b) Early biofilm treated with PMNs. (c) Mature biofilm treated without PMNs. (d) Mature biofilm treated with PMNs. Cells staining red are considered dead while cells staining green are viable cells.
Figure Legend Snippet: Effect of PMNs on biofilms of mucoid P. aeruginosa FRD1. Confocal laser scanning micrographs of P. aeruginosa FRD1 biofilms treated with PMNs. (a) Control group, early biofilm treated without PMNs. (b) Early biofilm treated with PMNs. (c) Mature biofilm treated without PMNs. (d) Mature biofilm treated with PMNs. Cells staining red are considered dead while cells staining green are viable cells.

Techniques Used: Staining

34) Product Images from "Polymorphonuclear Leukocytes or Hydrogen Peroxide Enhance Biofilm Development of Mucoid Pseudomonas aeruginosa"

Article Title: Polymorphonuclear Leukocytes or Hydrogen Peroxide Enhance Biofilm Development of Mucoid Pseudomonas aeruginosa

Journal: Mediators of Inflammation

doi: 10.1155/2018/8151362

Effect of PMNs and H 2 O 2 on viable cells of biofilms of mucoid P. aeruginosa FRD1. The error bars indicate standard deviations. Control: control group; PMNs: FRD1 biofilm group treated with PMNs; 1 mM H 2 O 2 : FRD1 biofilm group treated with 1 mM H 2 O 2 ; 2 mM H 2 O 2 : FRD1 biofilm group treated with 2 mM H 2 O 2 . (a, b) The numbers of viable cells in biofilms treated with PMNs are expressed in colony-forming units (CFUs). (c, d) The numbers of viable cells in biofilms treated with H 2 O 2 are expressed in colony-forming units (CFUs). Data are presented as the mean ± SD ( n = 6 in each treatment). ∗ P
Figure Legend Snippet: Effect of PMNs and H 2 O 2 on viable cells of biofilms of mucoid P. aeruginosa FRD1. The error bars indicate standard deviations. Control: control group; PMNs: FRD1 biofilm group treated with PMNs; 1 mM H 2 O 2 : FRD1 biofilm group treated with 1 mM H 2 O 2 ; 2 mM H 2 O 2 : FRD1 biofilm group treated with 2 mM H 2 O 2 . (a, b) The numbers of viable cells in biofilms treated with PMNs are expressed in colony-forming units (CFUs). (c, d) The numbers of viable cells in biofilms treated with H 2 O 2 are expressed in colony-forming units (CFUs). Data are presented as the mean ± SD ( n = 6 in each treatment). ∗ P

Techniques Used:

Effect of PMNs and H 2 O 2 on thickness of biofilms of mucoid P. aeruginosa FRD1. The error bars indicate standard deviations. Control: control group; PMNs: FRD1 biofilm group treated with PMNs; 1 mM H 2 O 2 : FRD1 biofilm group treated with 1 mM H 2 O 2 ; 2 mM H 2 O 2 : FRD1 biofilm group treated with 2 mM H 2 O 2 . (a, b) The thickness of biofilms treated with PMNs was determined using confocal laser-scanning microscopy. (c, d) The thickness of biofilms treated with H 2 O 2 was determined using confocal laser-scanning microscopy. Data are presented as the mean ± SD ( n = 6 in each treatment). ∗ P
Figure Legend Snippet: Effect of PMNs and H 2 O 2 on thickness of biofilms of mucoid P. aeruginosa FRD1. The error bars indicate standard deviations. Control: control group; PMNs: FRD1 biofilm group treated with PMNs; 1 mM H 2 O 2 : FRD1 biofilm group treated with 1 mM H 2 O 2 ; 2 mM H 2 O 2 : FRD1 biofilm group treated with 2 mM H 2 O 2 . (a, b) The thickness of biofilms treated with PMNs was determined using confocal laser-scanning microscopy. (c, d) The thickness of biofilms treated with H 2 O 2 was determined using confocal laser-scanning microscopy. Data are presented as the mean ± SD ( n = 6 in each treatment). ∗ P

Techniques Used: Confocal Laser Scanning Microscopy

Effect of nonactivated PMNs on viable cells of biofilms of mucoid P. aeruginosa FRD1. The numbers of viable cells in biofilms treated with PMNs are expressed in colony-forming units (CFUs). The error bars indicate standard deviations. BF: biofilm without PMNs; BF + nonactivated PMNs: biofilm treated with nonactivated PMNs. Data are presented as the mean ± SD ( n = 6 in each treatment).
Figure Legend Snippet: Effect of nonactivated PMNs on viable cells of biofilms of mucoid P. aeruginosa FRD1. The numbers of viable cells in biofilms treated with PMNs are expressed in colony-forming units (CFUs). The error bars indicate standard deviations. BF: biofilm without PMNs; BF + nonactivated PMNs: biofilm treated with nonactivated PMNs. Data are presented as the mean ± SD ( n = 6 in each treatment).

Techniques Used:

Effect of PMNs on G6PDH activity. G6PDH: glucose-6-phosphate dehydrogenase; Control: P. aeruginosa FRD1 biofilm grown for 24 hours or 48 hours without PMNs; PMN: P. aeruginosa FRD1 biofilm treated with PMNs for 24 hours or 48 hours. The activity of G6PDH did not differ significantly between the two groups. Data are presented as the mean ± SD ( n = 6 in each treatment).
Figure Legend Snippet: Effect of PMNs on G6PDH activity. G6PDH: glucose-6-phosphate dehydrogenase; Control: P. aeruginosa FRD1 biofilm grown for 24 hours or 48 hours without PMNs; PMN: P. aeruginosa FRD1 biofilm treated with PMNs for 24 hours or 48 hours. The activity of G6PDH did not differ significantly between the two groups. Data are presented as the mean ± SD ( n = 6 in each treatment).

Techniques Used: Activity Assay

Effect of PMNs on GMD activity. GMD: GDP-mannose dehydrogenase; Control: P. aeruginosa FRD1 biofilm grown for 24 hours or 48 hours without PMNs; PMN: P. aeruginosa FRD1 biofilm treated with PMNs for 24 hours or 48 hours. Activities of GMD in FRD1 biofilms treated with PMNs were significantly different from those in FRD1 biofilms without PMNs. Data are presented as the means ± SD ( n = 6 in each treatment). ∗ P
Figure Legend Snippet: Effect of PMNs on GMD activity. GMD: GDP-mannose dehydrogenase; Control: P. aeruginosa FRD1 biofilm grown for 24 hours or 48 hours without PMNs; PMN: P. aeruginosa FRD1 biofilm treated with PMNs for 24 hours or 48 hours. Activities of GMD in FRD1 biofilms treated with PMNs were significantly different from those in FRD1 biofilms without PMNs. Data are presented as the means ± SD ( n = 6 in each treatment). ∗ P

Techniques Used: Activity Assay

Effect of PMNs and H 2 O 2 on alginate content. The error bars indicate standard deviations. Control: control group; PMNs: FRD1 biofilm group treated with PMNs; 1 mM H 2 O 2 : FRD1 biofilm group treated with 1 mM H 2 O 2 ; 2 mM H 2 O 2 : FRD1 biofilm group treated with 2 mM H 2 O 2 . (a, b)The alginate content of P. aeruginosa FRD1 biofilms treated with PMNs. (c, d) The alginate content of P. aeruginosa FRD1 biofilms treated with H 2 O 2 . Data are presented as the mean ± SD ( n = 6 in each treatment). ∗ P
Figure Legend Snippet: Effect of PMNs and H 2 O 2 on alginate content. The error bars indicate standard deviations. Control: control group; PMNs: FRD1 biofilm group treated with PMNs; 1 mM H 2 O 2 : FRD1 biofilm group treated with 1 mM H 2 O 2 ; 2 mM H 2 O 2 : FRD1 biofilm group treated with 2 mM H 2 O 2 . (a, b)The alginate content of P. aeruginosa FRD1 biofilms treated with PMNs. (c, d) The alginate content of P. aeruginosa FRD1 biofilms treated with H 2 O 2 . Data are presented as the mean ± SD ( n = 6 in each treatment). ∗ P

Techniques Used:

Effects of PMNs and H 2 O 2 on the expression of genes involved in alginate biosynthesis. Control: control group; PMNs: FRD1 biofilm group treated with PMNs; 1 mM H 2 O 2 : FRD1 biofilm group treated with 1 mM H 2 O 2 ; 2 mM H 2 O 2 : FRD1 biofilm group treated with 2 mM H 2 O 2 . Bacteria were grown in Jensen's medium. RNA was extracted from the bacteria, and the relative mRNA levels of algD , algR , and algU were determined by RT-PCR. (a, b) The expression of genes involved in alginate biosynthesis after treatment with PMNs. (c, d) The expression of genes involved in alginate biosynthesis after treatment with H 2 O 2 . Data are presented as the mean ± SD ( n = 6 in each treatment). ∗ P
Figure Legend Snippet: Effects of PMNs and H 2 O 2 on the expression of genes involved in alginate biosynthesis. Control: control group; PMNs: FRD1 biofilm group treated with PMNs; 1 mM H 2 O 2 : FRD1 biofilm group treated with 1 mM H 2 O 2 ; 2 mM H 2 O 2 : FRD1 biofilm group treated with 2 mM H 2 O 2 . Bacteria were grown in Jensen's medium. RNA was extracted from the bacteria, and the relative mRNA levels of algD , algR , and algU were determined by RT-PCR. (a, b) The expression of genes involved in alginate biosynthesis after treatment with PMNs. (c, d) The expression of genes involved in alginate biosynthesis after treatment with H 2 O 2 . Data are presented as the mean ± SD ( n = 6 in each treatment). ∗ P

Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

Effect of PMA on viable cells of biofilms of mucoid P. aeruginosa FRD1. The numbers of viable cells in biofilms treated with PMA are expressed in colony-forming units (CFUs). The error bars indicate standard deviations. BF: biofilm without PMA; BF + PMA: biofilm treated with PMA (100 ng/ml). Data are presented as the mean ± SD ( n = 6 in each treatment).
Figure Legend Snippet: Effect of PMA on viable cells of biofilms of mucoid P. aeruginosa FRD1. The numbers of viable cells in biofilms treated with PMA are expressed in colony-forming units (CFUs). The error bars indicate standard deviations. BF: biofilm without PMA; BF + PMA: biofilm treated with PMA (100 ng/ml). Data are presented as the mean ± SD ( n = 6 in each treatment).

Techniques Used:

Effect of PMNs and H 2 O 2 on the adhesion of mucoid P. aeruginosa FRD1. The error bars indicate standard deviations. Control: control group; PMNs: FRD1 group treated with PMNs; 1 mM H 2 O 2 : FRD1 group treated with 1 mM H 2 O 2 ; 2 mM H 2 O 2 : FRD1 group treated with 2 mM H 2 O 2 . (a) Quantification of biofilm formation using crystal violet staining. Cells were grown in Jensen's medium for one day, in the presence of PMNs. PMNs increased the adhesion of P. aeruginosa FRD1. (b) The effect of PMNs on the number of viable cells is expressed in colony-forming units (CFUs). (c) Quantification of biofilm formation by crystal violet staining. H 2 O 2 increased the adhesion of P. aeruginosa FRD1. (d) The effect of H 2 O 2 on the number of viable cells is expressed in colony-forming units (CFUs). Data are presented as the mean ± SD ( n = 6 in each treatment). ∗ P
Figure Legend Snippet: Effect of PMNs and H 2 O 2 on the adhesion of mucoid P. aeruginosa FRD1. The error bars indicate standard deviations. Control: control group; PMNs: FRD1 group treated with PMNs; 1 mM H 2 O 2 : FRD1 group treated with 1 mM H 2 O 2 ; 2 mM H 2 O 2 : FRD1 group treated with 2 mM H 2 O 2 . (a) Quantification of biofilm formation using crystal violet staining. Cells were grown in Jensen's medium for one day, in the presence of PMNs. PMNs increased the adhesion of P. aeruginosa FRD1. (b) The effect of PMNs on the number of viable cells is expressed in colony-forming units (CFUs). (c) Quantification of biofilm formation by crystal violet staining. H 2 O 2 increased the adhesion of P. aeruginosa FRD1. (d) The effect of H 2 O 2 on the number of viable cells is expressed in colony-forming units (CFUs). Data are presented as the mean ± SD ( n = 6 in each treatment). ∗ P

Techniques Used: Staining

Effect of PMNs on biofilms of mucoid P. aeruginosa FRD1. Confocal laser scanning micrographs of P. aeruginosa FRD1 biofilms treated with PMNs. (a) Control group, early biofilm treated without PMNs. (b) Early biofilm treated with PMNs. (c) Mature biofilm treated without PMNs. (d) Mature biofilm treated with PMNs. Cells staining red are considered dead while cells staining green are viable cells.
Figure Legend Snippet: Effect of PMNs on biofilms of mucoid P. aeruginosa FRD1. Confocal laser scanning micrographs of P. aeruginosa FRD1 biofilms treated with PMNs. (a) Control group, early biofilm treated without PMNs. (b) Early biofilm treated with PMNs. (c) Mature biofilm treated without PMNs. (d) Mature biofilm treated with PMNs. Cells staining red are considered dead while cells staining green are viable cells.

Techniques Used: Staining

35) Product Images from "Dynamically Crosslinked Polymer Nanocomposites to Treat Multidrug-Resistant Bacterial Biofilms"

Article Title: Dynamically Crosslinked Polymer Nanocomposites to Treat Multidrug-Resistant Bacterial Biofilms

Journal: Nanoscale

doi: 10.1039/c8nr06348f

Viability of 1-day-old (a) P. aeruginosa (CD-1006), (b) E. coli (CD-2), (c) En. cloacae complex (CD-1412) and (d) S. aureus (CD-489) biofilms after 3 hour treatment with 5 wt% DCPN, PONI-GAT only, and Carvacrol oil only at different antimicrobial concentrations (mM)/(v/v %). The data are average of triplicates, and the error bars indicate the standard deviations.
Figure Legend Snippet: Viability of 1-day-old (a) P. aeruginosa (CD-1006), (b) E. coli (CD-2), (c) En. cloacae complex (CD-1412) and (d) S. aureus (CD-489) biofilms after 3 hour treatment with 5 wt% DCPN, PONI-GAT only, and Carvacrol oil only at different antimicrobial concentrations (mM)/(v/v %). The data are average of triplicates, and the error bars indicate the standard deviations.

Techniques Used:

Representative 3D views of confocal images stacks of RFP-expressing DH5-α E. coli biofilms, DiO-loaded DCPN and their overlay after treating the biofilm for 1 h with 5 wt% DiO-Loaded DCPN at 5 v/v% concentration in M9 media.
Figure Legend Snippet: Representative 3D views of confocal images stacks of RFP-expressing DH5-α E. coli biofilms, DiO-loaded DCPN and their overlay after treating the biofilm for 1 h with 5 wt% DiO-Loaded DCPN at 5 v/v% concentration in M9 media.

Techniques Used: Expressing, Concentration Assay

36) Product Images from "Role of de-N-acetylase PgaB from Aggregatibacter actinomycetemcomitans in exopolysaccharide export in biofilm mode of growth"

Article Title: Role of de-N-acetylase PgaB from Aggregatibacter actinomycetemcomitans in exopolysaccharide export in biofilm mode of growth

Journal: Molecular oral microbiology

doi: 10.1111/omi.12188

The role of the catalytic domain of PgaB in phenotypic variation and biofilm integrity. (a) Deletion of catalytic domain of PgaB (ΔNpgaB) results in a phenotype with attenuated rugged edges characteristic of IDH 781 but lacking the internal star
Figure Legend Snippet: The role of the catalytic domain of PgaB in phenotypic variation and biofilm integrity. (a) Deletion of catalytic domain of PgaB (ΔNpgaB) results in a phenotype with attenuated rugged edges characteristic of IDH 781 but lacking the internal star

Techniques Used:

Confocal scanning laser microscopic image of biofilms. (a) Biofilm growth was imaged at seven different locations and averaged for analysis. The scale bar is at 20 μm; (b) Cell viability for IDH 781 and ΔNpgaB strains at different time
Figure Legend Snippet: Confocal scanning laser microscopic image of biofilms. (a) Biofilm growth was imaged at seven different locations and averaged for analysis. The scale bar is at 20 μm; (b) Cell viability for IDH 781 and ΔNpgaB strains at different time

Techniques Used:

Immunofluorescence evaluation of biofilms for the production of PNAG. PNAG was visualized using the human mAb F598 and a secondary anti-human IgG conjugated to Alexa 488 (left panel). Images of the same field viewed by DAPI to stain DNA is shown in the
Figure Legend Snippet: Immunofluorescence evaluation of biofilms for the production of PNAG. PNAG was visualized using the human mAb F598 and a secondary anti-human IgG conjugated to Alexa 488 (left panel). Images of the same field viewed by DAPI to stain DNA is shown in the

Techniques Used: Immunofluorescence, Staining

37) Product Images from "Effects of Norspermidine on Dual‐Species Biofilms Composed of Streptococcus mutans and Streptococcus sanguinis"

Article Title: Effects of Norspermidine on Dual‐Species Biofilms Composed of Streptococcus mutans and Streptococcus sanguinis

Journal: BioMed Research International

doi: 10.1155/2019/1950790

Extracellular polysaccharides (EPS) in S. mutans and S. sanguinis dual-species biofilms. (a) EPS and bacterial staining of dual-species biofilm: (A–E) biofilm staining in the 0.0 mM norspermidine group, 7.0 mM norspermidine group, 3.5 mM norspermidine group, pH-adjusted (pH = 7.0) 7.0 mM norspermidine group, and pH-adjusted (pH = 7.0) 3.5 mM norspermidine group, respectively (bacteria were stained green and EPS was stained red, scale bar = 50 μ m); (b) EPS/bacteria ratio in dual-species biofilm according to coverage area; (c) water-insoluble glucans in biofilm of different groups. Data are presented as mean ± standard deviation, and values with dissimilar letters are significantly different from each other ( p
Figure Legend Snippet: Extracellular polysaccharides (EPS) in S. mutans and S. sanguinis dual-species biofilms. (a) EPS and bacterial staining of dual-species biofilm: (A–E) biofilm staining in the 0.0 mM norspermidine group, 7.0 mM norspermidine group, 3.5 mM norspermidine group, pH-adjusted (pH = 7.0) 7.0 mM norspermidine group, and pH-adjusted (pH = 7.0) 3.5 mM norspermidine group, respectively (bacteria were stained green and EPS was stained red, scale bar = 50 μ m); (b) EPS/bacteria ratio in dual-species biofilm according to coverage area; (c) water-insoluble glucans in biofilm of different groups. Data are presented as mean ± standard deviation, and values with dissimilar letters are significantly different from each other ( p

Techniques Used: Staining, Standard Deviation

SEM imaging, metabolic activity, and acid production of S. mutans and S. sanguinis dual-species biofilms. (a) SEM imaging of dual-species biofilms: (A–E) biofilm in the 0.0 mM norspermidine group, 7.0 mM norspermidine group, 3.5 mM norspermidine group, pH-adjusted (pH = 7.0) 7.0 mM norspermidine group, and pH-adjusted (pH = 7.0) 3.5 mM norspermidine group, respectively (5000x, scale bar = 20 μ m); (F–J) biofilm in the 0.0 mM norspermidine group, 7.0 mM norspermidine group, 3.5 mM norspermidine group, pH-adjusted (pH = 7.0) 7.0 mM norspermidine group, pH-adjusted (pH = 7.0) 3.5 mM norspermidine group, respectively (20000x, scale bar = 5 μ m); (b) metabolic activity of dual-species biofilm in different groups as revealed by MTT assay; (c) lactic acid production of dual-species biofilm revealed by lactic acid measurement. Data are presented as mean ± standard deviation, and values with dissimilar letters are significantly different from each other ( p
Figure Legend Snippet: SEM imaging, metabolic activity, and acid production of S. mutans and S. sanguinis dual-species biofilms. (a) SEM imaging of dual-species biofilms: (A–E) biofilm in the 0.0 mM norspermidine group, 7.0 mM norspermidine group, 3.5 mM norspermidine group, pH-adjusted (pH = 7.0) 7.0 mM norspermidine group, and pH-adjusted (pH = 7.0) 3.5 mM norspermidine group, respectively (5000x, scale bar = 20 μ m); (F–J) biofilm in the 0.0 mM norspermidine group, 7.0 mM norspermidine group, 3.5 mM norspermidine group, pH-adjusted (pH = 7.0) 7.0 mM norspermidine group, pH-adjusted (pH = 7.0) 3.5 mM norspermidine group, respectively (20000x, scale bar = 5 μ m); (b) metabolic activity of dual-species biofilm in different groups as revealed by MTT assay; (c) lactic acid production of dual-species biofilm revealed by lactic acid measurement. Data are presented as mean ± standard deviation, and values with dissimilar letters are significantly different from each other ( p

Techniques Used: Imaging, Activity Assay, MTT Assay, Standard Deviation

Single-species biofilm formation under the effects of norspermidine: (a) crystal violet staining of S. mutans and S. sanguinis biofilms under the effects of norspermidine; quantitative analysis of crystal violet staining of S. mutans biofilm (b) and S. sanguinis biofilm (c) under the effects of norspermidine. Data are presented as mean ± standard deviation, and values with dissimilar letters are significantly different from each other ( p
Figure Legend Snippet: Single-species biofilm formation under the effects of norspermidine: (a) crystal violet staining of S. mutans and S. sanguinis biofilms under the effects of norspermidine; quantitative analysis of crystal violet staining of S. mutans biofilm (b) and S. sanguinis biofilm (c) under the effects of norspermidine. Data are presented as mean ± standard deviation, and values with dissimilar letters are significantly different from each other ( p

Techniques Used: Staining, Standard Deviation

Composition of S. mutans and S. sanguinis dual-species biofilms as revealed by fluorescence in situ hybridization (FISH): (a)–(e) Dual-species biofilms in the 0.0 mM norspermidine group, 7.0 mM norspermidine group, 3.5 mM norspermidine group, pH-adjusted (pH = 7.0) 7.0 mM norspermidine group, and pH-adjusted (pH = 7.0) 3.5 mM norspermidine group, respectively ( S. mutans was stained green and S. sanguinis was stained red, scale bar = 50 μ m); (f) S. sanguinis ratio in dual-species biofilms according to coverage area. Data are presented as mean ± standard deviation, and values with dissimilar letters are significantly different from each other ( p
Figure Legend Snippet: Composition of S. mutans and S. sanguinis dual-species biofilms as revealed by fluorescence in situ hybridization (FISH): (a)–(e) Dual-species biofilms in the 0.0 mM norspermidine group, 7.0 mM norspermidine group, 3.5 mM norspermidine group, pH-adjusted (pH = 7.0) 7.0 mM norspermidine group, and pH-adjusted (pH = 7.0) 3.5 mM norspermidine group, respectively ( S. mutans was stained green and S. sanguinis was stained red, scale bar = 50 μ m); (f) S. sanguinis ratio in dual-species biofilms according to coverage area. Data are presented as mean ± standard deviation, and values with dissimilar letters are significantly different from each other ( p

Techniques Used: Fluorescence, In Situ Hybridization, Fluorescence In Situ Hybridization, Staining, Standard Deviation

38) Product Images from "The Type II Secretion System and Its Ubiquitous Lipoprotein Substrate, SslE, Are Required for Biofilm Formation and Virulence of Enteropathogenic Escherichia coli"

Article Title: The Type II Secretion System and Its Ubiquitous Lipoprotein Substrate, SslE, Are Required for Biofilm Formation and Virulence of Enteropathogenic Escherichia coli

Journal: Infection and Immunity

doi: 10.1128/IAI.06160-11

Proteomic and biofilm analysis of REPEC E22 and its derivatives. (A) SslE is secreted by the T2SS of REPEC strain E22. SslE (arrow) was detected in the supernatant of wild-type strain E22 (lane 1) and the trans -complemented mutants, E2348/69Δ gspD (pJP51) (lane 3) and E2348/69Δ gspK (pJP49), but not in the isogenic T2SS mutants, E2348/69Δ gspD (lane 2) and E2348/69Δ gspK (lane 4). Strains were grown in LB for 5 h. Proteins in the supernatants were precipitated in 10% trichloroacetic acid and washed in 25% acetone. Proteins were separated by SDS-PAGE using 4–12% bis-Tris NuPAGE gels (Invitrogen) and stained with Coomassie brilliant blue R-250. (B) Confocal laser scanning images of wild-type E22 and its isogenic mutants, E2348/69Δ gspD , E2348/69Δ gspK (T2SS mutants), and E2348/69Δ sslE , and the trans -complemented mutants, E2348/69Δ gspD (pJP51), E2348/69Δ gspK (pJP49), and E2348/69Δ sslE (pJP16). Shadow fluorescence projection images were rendered in Imaris. Bar, 50 μm. (C) Quantification of three-dimensional biofilm image stacks of wild-type E22 and its isogenic mutants, E2348/69Δ gspD , E2348/69Δ gspK (T2SS mutants), and E2348/69Δ sslE , these mutants complemented with empty vector, E2348/69Δ gspD (pACYC184), E2348/69Δ gspK (pACYC184), and E2348/69Δ sslE (pSU39), and trans -complemented mutants, E2348/69Δ gspD (pJP51), E2348/69Δ gspK (pJP49), and E2348/69Δ sslE (pJP16). T2SS and sslE mutants show significant defects in biofilm formation. Mean biomass was calculated by using COMSTAT analysis of at least eight random image stacks from each strain. Values represent means (±SEM) of three independent experiments expressed as a percentage of the wild type (***, P
Figure Legend Snippet: Proteomic and biofilm analysis of REPEC E22 and its derivatives. (A) SslE is secreted by the T2SS of REPEC strain E22. SslE (arrow) was detected in the supernatant of wild-type strain E22 (lane 1) and the trans -complemented mutants, E2348/69Δ gspD (pJP51) (lane 3) and E2348/69Δ gspK (pJP49), but not in the isogenic T2SS mutants, E2348/69Δ gspD (lane 2) and E2348/69Δ gspK (lane 4). Strains were grown in LB for 5 h. Proteins in the supernatants were precipitated in 10% trichloroacetic acid and washed in 25% acetone. Proteins were separated by SDS-PAGE using 4–12% bis-Tris NuPAGE gels (Invitrogen) and stained with Coomassie brilliant blue R-250. (B) Confocal laser scanning images of wild-type E22 and its isogenic mutants, E2348/69Δ gspD , E2348/69Δ gspK (T2SS mutants), and E2348/69Δ sslE , and the trans -complemented mutants, E2348/69Δ gspD (pJP51), E2348/69Δ gspK (pJP49), and E2348/69Δ sslE (pJP16). Shadow fluorescence projection images were rendered in Imaris. Bar, 50 μm. (C) Quantification of three-dimensional biofilm image stacks of wild-type E22 and its isogenic mutants, E2348/69Δ gspD , E2348/69Δ gspK (T2SS mutants), and E2348/69Δ sslE , these mutants complemented with empty vector, E2348/69Δ gspD (pACYC184), E2348/69Δ gspK (pACYC184), and E2348/69Δ sslE (pSU39), and trans -complemented mutants, E2348/69Δ gspD (pJP51), E2348/69Δ gspK (pJP49), and E2348/69Δ sslE (pJP16). T2SS and sslE mutants show significant defects in biofilm formation. Mean biomass was calculated by using COMSTAT analysis of at least eight random image stacks from each strain. Values represent means (±SEM) of three independent experiments expressed as a percentage of the wild type (***, P

Techniques Used: SDS Page, Staining, Fluorescence, Plasmid Preparation

Analysis of biofilm formation by EPEC E2348/69 and its derivatives. (A) Confocal laser scanning images of wild-type E2348/69 and its isogenic mutants, E2348/69Δ gspD (T2SS mutant) and E2348/69Δ sslE , and trans -complemented mutants, E2348/69Δ gspD (pJP52) and E2348/69Δ sslE (pJP15). Shadow fluorescence projection images are rendered in Imaris. Bar, 50 μm. (B) Quantification of three-dimensional biofilm image stacks of wild-type E2348/69 and its isogenic mutants, E2348/69Δ gspD , and E2348/69Δ sslE , trans -complemented mutants, E2348/69Δ gspD (pJP52) and E2348/69Δ sslE (pJP15), and these mutants complemented with empty vector, E2348/69Δ gspD (pACYC184) and E2348/69Δ sslE (pSU39). T2SS and sslE ) of at least eight random image stacks from each strain. Values represent the means (±SEM) of three independent experiments expressed as a percentage of the wild type (***, P
Figure Legend Snippet: Analysis of biofilm formation by EPEC E2348/69 and its derivatives. (A) Confocal laser scanning images of wild-type E2348/69 and its isogenic mutants, E2348/69Δ gspD (T2SS mutant) and E2348/69Δ sslE , and trans -complemented mutants, E2348/69Δ gspD (pJP52) and E2348/69Δ sslE (pJP15). Shadow fluorescence projection images are rendered in Imaris. Bar, 50 μm. (B) Quantification of three-dimensional biofilm image stacks of wild-type E2348/69 and its isogenic mutants, E2348/69Δ gspD , and E2348/69Δ sslE , trans -complemented mutants, E2348/69Δ gspD (pJP52) and E2348/69Δ sslE (pJP15), and these mutants complemented with empty vector, E2348/69Δ gspD (pACYC184) and E2348/69Δ sslE (pSU39). T2SS and sslE ) of at least eight random image stacks from each strain. Values represent the means (±SEM) of three independent experiments expressed as a percentage of the wild type (***, P

Techniques Used: Mutagenesis, Fluorescence, Plasmid Preparation

39) Product Images from "Biofilm Formation by Pneumocystis spp. ▿ spp. ▿ †"

Article Title: Biofilm Formation by Pneumocystis spp. ▿ spp. ▿ †

Journal:

doi: 10.1128/EC.00202-08

Process of biofilm formation monitored by phase microscopy. Phase microscopy of wet mounts (unstained) of P. carinii biofilms is shown. (A) Cluster of P. carinii after 4 days in the Millicell-CM insert system. Arrows indicate focal points of refractility.
Figure Legend Snippet: Process of biofilm formation monitored by phase microscopy. Phase microscopy of wet mounts (unstained) of P. carinii biofilms is shown. (A) Cluster of P. carinii after 4 days in the Millicell-CM insert system. Arrows indicate focal points of refractility.

Techniques Used: Microscopy

Autofluorescence of a P. carinii biofilm cluster. (A) Differential interference contrast image of a cluster with anastomosed extension from a 7-day-old biofilm; (B) same cluster showing autofluorescence at 488-nm excitation. Bar, 10 μm.
Figure Legend Snippet: Autofluorescence of a P. carinii biofilm cluster. (A) Differential interference contrast image of a cluster with anastomosed extension from a 7-day-old biofilm; (B) same cluster showing autofluorescence at 488-nm excitation. Bar, 10 μm.

Techniques Used:

Macroscopic biofilm formation by P. carinii . (A) Duplicate wells showing biofilm formation after 1, 5, and 10 days on a Lab Tek II chamber slide. (B) PTFE inserts (12 mm) without inocula (left) and after 14 days of biofilm formation (right). Photographs
Figure Legend Snippet: Macroscopic biofilm formation by P. carinii . (A) Duplicate wells showing biofilm formation after 1, 5, and 10 days on a Lab Tek II chamber slide. (B) PTFE inserts (12 mm) without inocula (left) and after 14 days of biofilm formation (right). Photographs

Techniques Used:

Merged orthogonal images of P. carinii biofilms with FUN-1 and ConA-Alexafluor. After 7 days of culture in Lab Tek II chambers, FUN-1 and ConA-Alexafluor 488 were added to the P. carinii culture medium and cells were incubated, washed, and mounted in
Figure Legend Snippet: Merged orthogonal images of P. carinii biofilms with FUN-1 and ConA-Alexafluor. After 7 days of culture in Lab Tek II chambers, FUN-1 and ConA-Alexafluor 488 were added to the P. carinii culture medium and cells were incubated, washed, and mounted in

Techniques Used: Incubation

MSGs are abundant on the surface of P. carinii biofilms. Staining reactivity of monoclonal antibody RA-E7, directed to a protein epitope of the MSG family of antigens, with a cluster from a 7-day P. carinii biofilm is shown. Bar, 10 μm.
Figure Legend Snippet: MSGs are abundant on the surface of P. carinii biofilms. Staining reactivity of monoclonal antibody RA-E7, directed to a protein epitope of the MSG family of antigens, with a cluster from a 7-day P. carinii biofilm is shown. Bar, 10 μm.

Techniques Used: Staining

(1,3)-β- d -glucan content in P. carinii biofilms. Duplicate Millicell-CM inserts were harvested over a 3-week period and the (1,3)-β- d -glucan content measured using the Glucatell assay kit as described in Materials and Methods. Day 0 refers
Figure Legend Snippet: (1,3)-β- d -glucan content in P. carinii biofilms. Duplicate Millicell-CM inserts were harvested over a 3-week period and the (1,3)-β- d -glucan content measured using the Glucatell assay kit as described in Materials and Methods. Day 0 refers

Techniques Used:

Inoculation of passaged biofilms into P. carinii -naïve rats.
Figure Legend Snippet: Inoculation of passaged biofilms into P. carinii -naïve rats.

Techniques Used:

ATP content of P. carinii biofilms exposed to farnesol. Farnesol (100 μM) was added to the inserts with the inocula. Biofilms were fed with medium containing the farnesol on a daily basis. Bars reflect the percent ATP content compared to untreated
Figure Legend Snippet: ATP content of P. carinii biofilms exposed to farnesol. Farnesol (100 μM) was added to the inserts with the inocula. Biofilms were fed with medium containing the farnesol on a daily basis. Bars reflect the percent ATP content compared to untreated

Techniques Used:

Temporal development of P. carinii biofilms on inserts. Organisms were inoculated onto cell culture insert membranes at a density of 2 × 10 7 /insert, placed in multiwell plates, allowed to adhere for 24 h, washed with PBS, and placed back in the
Figure Legend Snippet: Temporal development of P. carinii biofilms on inserts. Organisms were inoculated onto cell culture insert membranes at a density of 2 × 10 7 /insert, placed in multiwell plates, allowed to adhere for 24 h, washed with PBS, and placed back in the

Techniques Used: Cell Culture

Biofilm formation by P. murina and P. carinii with highly refractile large clusters. (A and B) Large clusters of P. murina from 10-day-old biofilms viewed unstained under phase-contrast microscopy. Note that the clusters spanned well beyond the microscopic
Figure Legend Snippet: Biofilm formation by P. murina and P. carinii with highly refractile large clusters. (A and B) Large clusters of P. murina from 10-day-old biofilms viewed unstained under phase-contrast microscopy. Note that the clusters spanned well beyond the microscopic

Techniques Used: Microscopy

The morphology of Pneumocystis changes dramatically during biofilm formation. (A) P. carinii from the supernatant of a 3-day-old standard short-term culture stained with Hema3, illustrating the differences in morphology from the biofilm structures. (B
Figure Legend Snippet: The morphology of Pneumocystis changes dramatically during biofilm formation. (A) P. carinii from the supernatant of a 3-day-old standard short-term culture stained with Hema3, illustrating the differences in morphology from the biofilm structures. (B

Techniques Used: Staining

40) Product Images from "Effects of Norspermidine on Dual‐Species Biofilms Composed of Streptococcus mutans and Streptococcus sanguinis"

Article Title: Effects of Norspermidine on Dual‐Species Biofilms Composed of Streptococcus mutans and Streptococcus sanguinis

Journal: BioMed Research International

doi: 10.1155/2019/1950790

Extracellular polysaccharides (EPS) in S. mutans and S. sanguinis dual-species biofilms. (a) EPS and bacterial staining of dual-species biofilm: (A–E) biofilm staining in the 0.0 mM norspermidine group, 7.0 mM norspermidine group, 3.5 mM norspermidine group, pH-adjusted (pH = 7.0) 7.0 mM norspermidine group, and pH-adjusted (pH = 7.0) 3.5 mM norspermidine group, respectively (bacteria were stained green and EPS was stained red, scale bar = 50 μ m); (b) EPS/bacteria ratio in dual-species biofilm according to coverage area; (c) water-insoluble glucans in biofilm of different groups. Data are presented as mean ± standard deviation, and values with dissimilar letters are significantly different from each other ( p
Figure Legend Snippet: Extracellular polysaccharides (EPS) in S. mutans and S. sanguinis dual-species biofilms. (a) EPS and bacterial staining of dual-species biofilm: (A–E) biofilm staining in the 0.0 mM norspermidine group, 7.0 mM norspermidine group, 3.5 mM norspermidine group, pH-adjusted (pH = 7.0) 7.0 mM norspermidine group, and pH-adjusted (pH = 7.0) 3.5 mM norspermidine group, respectively (bacteria were stained green and EPS was stained red, scale bar = 50 μ m); (b) EPS/bacteria ratio in dual-species biofilm according to coverage area; (c) water-insoluble glucans in biofilm of different groups. Data are presented as mean ± standard deviation, and values with dissimilar letters are significantly different from each other ( p

Techniques Used: Staining, Standard Deviation

SEM imaging, metabolic activity, and acid production of S. mutans and S. sanguinis dual-species biofilms. (a) SEM imaging of dual-species biofilms: (A–E) biofilm in the 0.0 mM norspermidine group, 7.0 mM norspermidine group, 3.5 mM norspermidine group, pH-adjusted (pH = 7.0) 7.0 mM norspermidine group, and pH-adjusted (pH = 7.0) 3.5 mM norspermidine group, respectively (5000x, scale bar = 20 μ m); (F–J) biofilm in the 0.0 mM norspermidine group, 7.0 mM norspermidine group, 3.5 mM norspermidine group, pH-adjusted (pH = 7.0) 7.0 mM norspermidine group, pH-adjusted (pH = 7.0) 3.5 mM norspermidine group, respectively (20000x, scale bar = 5 μ m); (b) metabolic activity of dual-species biofilm in different groups as revealed by MTT assay; (c) lactic acid production of dual-species biofilm revealed by lactic acid measurement. Data are presented as mean ± standard deviation, and values with dissimilar letters are significantly different from each other ( p
Figure Legend Snippet: SEM imaging, metabolic activity, and acid production of S. mutans and S. sanguinis dual-species biofilms. (a) SEM imaging of dual-species biofilms: (A–E) biofilm in the 0.0 mM norspermidine group, 7.0 mM norspermidine group, 3.5 mM norspermidine group, pH-adjusted (pH = 7.0) 7.0 mM norspermidine group, and pH-adjusted (pH = 7.0) 3.5 mM norspermidine group, respectively (5000x, scale bar = 20 μ m); (F–J) biofilm in the 0.0 mM norspermidine group, 7.0 mM norspermidine group, 3.5 mM norspermidine group, pH-adjusted (pH = 7.0) 7.0 mM norspermidine group, pH-adjusted (pH = 7.0) 3.5 mM norspermidine group, respectively (20000x, scale bar = 5 μ m); (b) metabolic activity of dual-species biofilm in different groups as revealed by MTT assay; (c) lactic acid production of dual-species biofilm revealed by lactic acid measurement. Data are presented as mean ± standard deviation, and values with dissimilar letters are significantly different from each other ( p

Techniques Used: Imaging, Activity Assay, MTT Assay, Standard Deviation

Single-species biofilm formation under the effects of norspermidine: (a) crystal violet staining of S. mutans and S. sanguinis biofilms under the effects of norspermidine; quantitative analysis of crystal violet staining of S. mutans biofilm (b) and S. sanguinis biofilm (c) under the effects of norspermidine. Data are presented as mean ± standard deviation, and values with dissimilar letters are significantly different from each other ( p
Figure Legend Snippet: Single-species biofilm formation under the effects of norspermidine: (a) crystal violet staining of S. mutans and S. sanguinis biofilms under the effects of norspermidine; quantitative analysis of crystal violet staining of S. mutans biofilm (b) and S. sanguinis biofilm (c) under the effects of norspermidine. Data are presented as mean ± standard deviation, and values with dissimilar letters are significantly different from each other ( p

Techniques Used: Staining, Standard Deviation

Composition of S. mutans and S. sanguinis dual-species biofilms as revealed by fluorescence in situ hybridization (FISH): (a)–(e) Dual-species biofilms in the 0.0 mM norspermidine group, 7.0 mM norspermidine group, 3.5 mM norspermidine group, pH-adjusted (pH = 7.0) 7.0 mM norspermidine group, and pH-adjusted (pH = 7.0) 3.5 mM norspermidine group, respectively ( S. mutans was stained green and S. sanguinis was stained red, scale bar = 50 μ m); (f) S. sanguinis ratio in dual-species biofilms according to coverage area. Data are presented as mean ± standard deviation, and values with dissimilar letters are significantly different from each other ( p
Figure Legend Snippet: Composition of S. mutans and S. sanguinis dual-species biofilms as revealed by fluorescence in situ hybridization (FISH): (a)–(e) Dual-species biofilms in the 0.0 mM norspermidine group, 7.0 mM norspermidine group, 3.5 mM norspermidine group, pH-adjusted (pH = 7.0) 7.0 mM norspermidine group, and pH-adjusted (pH = 7.0) 3.5 mM norspermidine group, respectively ( S. mutans was stained green and S. sanguinis was stained red, scale bar = 50 μ m); (f) S. sanguinis ratio in dual-species biofilms according to coverage area. Data are presented as mean ± standard deviation, and values with dissimilar letters are significantly different from each other ( p

Techniques Used: Fluorescence, In Situ Hybridization, Fluorescence In Situ Hybridization, Staining, Standard Deviation

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Fluorescence:

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Article Title: Citrullination mediated by PPAD constrains biofilm formation in P. gingivalis strain 381
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Laser-Scanning Microscopy:

Article Title: Discovery of new diketopiperazines inhibiting Burkholderia cenocepacia quorum sensing in vitro and in vivo
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Microscopy:

Article Title: Finding a Missing Gene: EFG1 Regulates Morphogenesis in Candida tropicalis
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Article Title: α-Chymotrypsin Immobilized on a Low-Density Polyethylene Surface Successfully Weakens Escherichia coli Biofilm Formation
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Article Title: Citrullination mediated by PPAD constrains biofilm formation in P. gingivalis strain 381
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Article Title: The LuxS/AI-2 Quorum-Sensing System of Streptococcus pneumoniae Is Required to Cause Disease, and to Regulate Virulence- and Metabolism-Related Genes in a Rat Model of Middle Ear Infection
Article Snippet: .. After washing with PBS, the stained biofilms were examined using a Nikon A1 confocal microscope (Nikon Instruments, Inc., NY, USA) with fluorescein (green) and Texas red (red) band-pass filter sets. .. Evaluation of the in vivo colonization capability of the S. pneumoniae D39 wild-type and D39ΔluxS The in vivo colonization capability of the D39 wild-type and D39ΔluxS strains was evaluated using a rat model of OM (Yadav et al., , ).

Incubation:

Article Title: Discovery of new diketopiperazines inhibiting Burkholderia cenocepacia quorum sensing in vitro and in vivo
Article Snippet: .. These plates were incubated in the dark for 15 min at room temperature and the biofilm was visualized with a Nikon C1 confocal laser scanning microscope (Nikon Benelux, Brussels, Belgium) as previously described . ..

Confocal Laser Scanning Microscopy:

Article Title: Synergistic Antifungal Effect of Amphotericin B-Loaded Poly(Lactic-Co-Glycolic Acid) Nanoparticles and Ultrasound against Candida albicans Biofilms
Article Snippet: .. The changes in the architecture of biofilms and living and dead fungi in the biofilm after different treatments were observed by confocal laser scanning microscopy (CLSM) (A1+R; Nikon, Tokyo, Japan). shows viable cells (green fluorescence) and dead cells (red fluorescence) in the bottom biofilm layer of three-dimensional (3-D) reconstructed images. .. The control group was dominated by green fluorescence, which showed dense growth of live fungal cells.

Staining:

Article Title: Finding a Missing Gene: EFG1 Regulates Morphogenesis in Candida tropicalis
Article Snippet: .. Biofilms formed on the squares were stained for 1 hr with 50 mg/ml of concanavalin A Alexa Fluor 594 conjugate and visualized on a confocal microscope using a 40×/0.80W Nikon objective. ..

Article Title: Citrullination mediated by PPAD constrains biofilm formation in P. gingivalis strain 381
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Software:

Article Title: Verticalization of bacterial biofilms
Article Snippet: .. For biofilm clusters grown in the presence of A22 or Cefalexin, cell lengths were manually measured in the bottom cell layers of the biofilms using the Nikon Element software. .. To define the biofilm shape parameters in the coarse-grained images, the bottom cell layers of the biofilms were first identified by finding the brightest z -cross section, according to the total fluorescence intensity.

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    Nikon biofilm clusters
    Mechanics of cell reorientation in modeled biofilms, ( a-b ) Properties of individual cells at the time t r of reorientation, defined as the time of the peak of total force on the cell prior to it becoming vertical. Analyses are shown for all reorientation events among different biofilms simulated for a range of initial cell lengths ℓ 0 ( a ) Distributions of reorientation “surface pressure” p r , defined as the total contact force in the xy plane acting on a cell at time t r , normalized by the cell’s perimeter, versus cell cylinder length ℓ . The white dashed curve shows the average reorientation surface pressure ⟨ p r ⟩ as a function of ℓ . The magenta dashed curve shows the threshold surface pressure p t from linear stability analysis for a modeled cell under uniform pressure, depicted schematically in the inset, ( b ) Distributions of the logarithm of reorientation torque τ r , defined as the magnitude of the torque on a cell due to cell-cell contact forces in the z direction at time t r , for different cell cylinder lengths ℓ . The white dashed curve shows the average values ⟨log τ r ⟩ as a function of ℓ . The orange dashed curve shows the scaling τ t ~ ℓ 2 of the threshold torque for peeling from linear stability analysis for a modeled cell, depicted schematically in the inset, ( c ) Mean reorientation length ⟨ ℓ r ⟩ (red), defined as the average value of cell length at t r , and mean cell cylinder length ⟨ ℓ ⟩ (gray), defined as the average length of all horizontal cells over all times of <t>biofilm</t> growth, averaged over ten simulated biofilms, each with initial cell cylinder length ℓ 0 , plotted versus ℓ 0 . The inset shows the distribution of reorientation lengths (red) and horizontal surface-cell lengths (gray) for ℓ 0 = 1 μm. ( d ) Mean avalanche size ⟨ N ⟩, defined as the average size of a cluster of reorienting cells that are proximal in space and time ( Supplementary Figs. 8 - 10 ), versus initial cell length ℓ 0 for the experimental biofilm (red triangle) and the modeled biofilm (red circles). Open gray triangle and circles indicate the corresponding mean avalanche sizes for a null model. Inset shows a side view of cell configurations in the xy plane at times t r for all reorientation events in a simulated biofilm with ℓ 0 = 2.5 μm. Reorientation events are colored alike if they belong to the same avalanche. Scale bars: 10 μm and 1 hour.
    Biofilm Clusters, supplied by Nikon, used in various techniques. Bioz Stars score: 92/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Characterization of the phenotype of BCR1 -phosphorylation mutants in <t>biofilm</t> formation in vitro . BCR1 phosphomimetic ( bcr1 T191E / bcr1 Δ, JC1094; bcr1 S556E / bcr1 Δ, JC1092; bcr1EE / bcr1 Δ, JC1180) and phosphodefective ( bcr1 T191A / bcr1 Δ, JC1093; bcr1 S556A / bcr1 Δ, JC1088; bcr1AA / bcr1 Δ, JC1178) mutants were grown in biofilm-inducing conditions for 40 hours using microfermentors. BCR1/bcr1 Δ (WT, JC1089) and bcr1 Δ (JC1081) strains were used as controls. (A) Quantification of adherent cells. (B) Biofilm dry mass determination. (C) Biofilms grown using silicon squares models were stained with calcofluor white for CSLM visualization.
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    Mechanics of cell reorientation in modeled biofilms, ( a-b ) Properties of individual cells at the time t r of reorientation, defined as the time of the peak of total force on the cell prior to it becoming vertical. Analyses are shown for all reorientation events among different biofilms simulated for a range of initial cell lengths ℓ 0 ( a ) Distributions of reorientation “surface pressure” p r , defined as the total contact force in the xy plane acting on a cell at time t r , normalized by the cell’s perimeter, versus cell cylinder length ℓ . The white dashed curve shows the average reorientation surface pressure ⟨ p r ⟩ as a function of ℓ . The magenta dashed curve shows the threshold surface pressure p t from linear stability analysis for a modeled cell under uniform pressure, depicted schematically in the inset, ( b ) Distributions of the logarithm of reorientation torque τ r , defined as the magnitude of the torque on a cell due to cell-cell contact forces in the z direction at time t r , for different cell cylinder lengths ℓ . The white dashed curve shows the average values ⟨log τ r ⟩ as a function of ℓ . The orange dashed curve shows the scaling τ t ~ ℓ 2 of the threshold torque for peeling from linear stability analysis for a modeled cell, depicted schematically in the inset, ( c ) Mean reorientation length ⟨ ℓ r ⟩ (red), defined as the average value of cell length at t r , and mean cell cylinder length ⟨ ℓ ⟩ (gray), defined as the average length of all horizontal cells over all times of biofilm growth, averaged over ten simulated biofilms, each with initial cell cylinder length ℓ 0 , plotted versus ℓ 0 . The inset shows the distribution of reorientation lengths (red) and horizontal surface-cell lengths (gray) for ℓ 0 = 1 μm. ( d ) Mean avalanche size ⟨ N ⟩, defined as the average size of a cluster of reorienting cells that are proximal in space and time ( Supplementary Figs. 8 - 10 ), versus initial cell length ℓ 0 for the experimental biofilm (red triangle) and the modeled biofilm (red circles). Open gray triangle and circles indicate the corresponding mean avalanche sizes for a null model. Inset shows a side view of cell configurations in the xy plane at times t r for all reorientation events in a simulated biofilm with ℓ 0 = 2.5 μm. Reorientation events are colored alike if they belong to the same avalanche. Scale bars: 10 μm and 1 hour.

    Journal: Nature physics

    Article Title: Verticalization of bacterial biofilms

    doi: 10.1038/s41567-018-0170-4

    Figure Lengend Snippet: Mechanics of cell reorientation in modeled biofilms, ( a-b ) Properties of individual cells at the time t r of reorientation, defined as the time of the peak of total force on the cell prior to it becoming vertical. Analyses are shown for all reorientation events among different biofilms simulated for a range of initial cell lengths ℓ 0 ( a ) Distributions of reorientation “surface pressure” p r , defined as the total contact force in the xy plane acting on a cell at time t r , normalized by the cell’s perimeter, versus cell cylinder length ℓ . The white dashed curve shows the average reorientation surface pressure ⟨ p r ⟩ as a function of ℓ . The magenta dashed curve shows the threshold surface pressure p t from linear stability analysis for a modeled cell under uniform pressure, depicted schematically in the inset, ( b ) Distributions of the logarithm of reorientation torque τ r , defined as the magnitude of the torque on a cell due to cell-cell contact forces in the z direction at time t r , for different cell cylinder lengths ℓ . The white dashed curve shows the average values ⟨log τ r ⟩ as a function of ℓ . The orange dashed curve shows the scaling τ t ~ ℓ 2 of the threshold torque for peeling from linear stability analysis for a modeled cell, depicted schematically in the inset, ( c ) Mean reorientation length ⟨ ℓ r ⟩ (red), defined as the average value of cell length at t r , and mean cell cylinder length ⟨ ℓ ⟩ (gray), defined as the average length of all horizontal cells over all times of biofilm growth, averaged over ten simulated biofilms, each with initial cell cylinder length ℓ 0 , plotted versus ℓ 0 . The inset shows the distribution of reorientation lengths (red) and horizontal surface-cell lengths (gray) for ℓ 0 = 1 μm. ( d ) Mean avalanche size ⟨ N ⟩, defined as the average size of a cluster of reorienting cells that are proximal in space and time ( Supplementary Figs. 8 - 10 ), versus initial cell length ℓ 0 for the experimental biofilm (red triangle) and the modeled biofilm (red circles). Open gray triangle and circles indicate the corresponding mean avalanche sizes for a null model. Inset shows a side view of cell configurations in the xy plane at times t r for all reorientation events in a simulated biofilm with ℓ 0 = 2.5 μm. Reorientation events are colored alike if they belong to the same avalanche. Scale bars: 10 μm and 1 hour.

    Article Snippet: For biofilm clusters grown in the presence of A22 or Cefalexin, cell lengths were manually measured in the bottom cell layers of the biofilms using the Nikon Element software.

    Techniques:

    Development of experimental and modeled biofilms. ( a , b ) Top-down and perspective visualizations of the surface layer of ( a ) experimental and ( b ) modeled biofilms, showing positions and orientations of horizontal (blue) and vertical (red) surface-adhered cells as spherocylinders of radius R = 0.8 μm, with the surface shown at height z = 0 μm (brown). Cells with n z

    Journal: Nature physics

    Article Title: Verticalization of bacterial biofilms

    doi: 10.1038/s41567-018-0170-4

    Figure Lengend Snippet: Development of experimental and modeled biofilms. ( a , b ) Top-down and perspective visualizations of the surface layer of ( a ) experimental and ( b ) modeled biofilms, showing positions and orientations of horizontal (blue) and vertical (red) surface-adhered cells as spherocylinders of radius R = 0.8 μm, with the surface shown at height z = 0 μm (brown). Cells with n z

    Article Snippet: For biofilm clusters grown in the presence of A22 or Cefalexin, cell lengths were manually measured in the bottom cell layers of the biofilms using the Nikon Element software.

    Techniques:

    Two-component fluid model for verticalizing cells in biofilms. ( a ) Schematic illustration of the two-component continuum model. Horizontal cells (blue) and vertical cells (red) are modeled, respectively, by densities ρ h and ρ v in two spatial dimensions. The total cell density ρ ~ tot is defined as ρ h + ξρ ν , where ξ is the ratio of vertical to horizontal cell footprints. ( b ) Radial densities ρ of vertical cells ( ρ v , red), horizontal cells ( ρ h , blue), and total density ( ρ ~ tot , black), versus shifted radial coordinate r ~ , defined as the radial position relative to the boundary between the mixed interior and the horizontal cell periphery. Results are shown for the continuum model (left; radial cell density in units of μm −2 ), the experimental biofilm (middle; radial cell density in each μm-sized bin averaged over an observation window of 50 minutes), and the agent-based model biofilm (right; radial cell density in each μm-sized bin averaged for ten biofilms over an observation window of 6 minutes). For the continuum model and the agent-based model biofilms the parameters were chosen to match those obtained from the experiment ( Supplementary Figs. 12 - 13 ). Inset in the left-most panel shows the fraction of vertical cells in the continuum model at a given radius from the biofilm center (gray regions contain no cells, color scale is the same as in Fig. 1 ).

    Journal: Nature physics

    Article Title: Verticalization of bacterial biofilms

    doi: 10.1038/s41567-018-0170-4

    Figure Lengend Snippet: Two-component fluid model for verticalizing cells in biofilms. ( a ) Schematic illustration of the two-component continuum model. Horizontal cells (blue) and vertical cells (red) are modeled, respectively, by densities ρ h and ρ v in two spatial dimensions. The total cell density ρ ~ tot is defined as ρ h + ξρ ν , where ξ is the ratio of vertical to horizontal cell footprints. ( b ) Radial densities ρ of vertical cells ( ρ v , red), horizontal cells ( ρ h , blue), and total density ( ρ ~ tot , black), versus shifted radial coordinate r ~ , defined as the radial position relative to the boundary between the mixed interior and the horizontal cell periphery. Results are shown for the continuum model (left; radial cell density in units of μm −2 ), the experimental biofilm (middle; radial cell density in each μm-sized bin averaged over an observation window of 50 minutes), and the agent-based model biofilm (right; radial cell density in each μm-sized bin averaged for ten biofilms over an observation window of 6 minutes). For the continuum model and the agent-based model biofilms the parameters were chosen to match those obtained from the experiment ( Supplementary Figs. 12 - 13 ). Inset in the left-most panel shows the fraction of vertical cells in the continuum model at a given radius from the biofilm center (gray regions contain no cells, color scale is the same as in Fig. 1 ).

    Article Snippet: For biofilm clusters grown in the presence of A22 or Cefalexin, cell lengths were manually measured in the bottom cell layers of the biofilms using the Nikon Element software.

    Techniques:

    Global morphological properties of experimental and modeled biofilms, ( a ) Top-down (upper row) and side views (lower row) of experimental biofilms grown with 0.4 μg/mL A22 (magenta), without treatment (yellow), and with 4 μg/mL Cefalexin (cyan), following overnight growth (upper row) and 7 hours after inoculation (lower row). Scale bar: 10 μm. Insets show magnifications of 10 μm 2 -sized regions of top-down views taken from the peripheries of biofilms. ( b ) Expansion speed c *, defined as the speed of the biofilm edge along the surface, versus the initial cell cylinder length ℓ 0 for experimental biofilms (A22, magenta; no treatment, yellow; Cefalexin, cyan), agent-based model biofilms (black circles), and continuum model (dashed black curve). Expansion velocities were determined from a linear fit of the basal radius R B of the biofilm versus time, where R B is defined at each time point as the radius of a circle with area equal to that of the biofilm base. For experimental biofilms, the boundary was extracted from the normalized fluorescence data (see Methods for details). For each treatment, the vertical error bars show the standard error of the mean of the expansion speed and the horizontal error bars bound the measured initial cell cylinder length ( Supplementary Fig. 1 ). Inset: model cells with lengths and radii corresponding to the averages for different treatments, ( c ) Biofilm aspect ratio H/R B for experimental biofilms grown under different treatments, where the biofilm height is defined as H = 3 V ∕ 2 R B 2 , the height of a semi-ellipsoid with a circular base of radius R B and volume V equal to that of the biofilm. Error bars show the standard error of the mean. Inset: overlay of biofilm outlines from bottom row of panel ( a ). Color designations and treatments same as in panel ( a ).

    Journal: Nature physics

    Article Title: Verticalization of bacterial biofilms

    doi: 10.1038/s41567-018-0170-4

    Figure Lengend Snippet: Global morphological properties of experimental and modeled biofilms, ( a ) Top-down (upper row) and side views (lower row) of experimental biofilms grown with 0.4 μg/mL A22 (magenta), without treatment (yellow), and with 4 μg/mL Cefalexin (cyan), following overnight growth (upper row) and 7 hours after inoculation (lower row). Scale bar: 10 μm. Insets show magnifications of 10 μm 2 -sized regions of top-down views taken from the peripheries of biofilms. ( b ) Expansion speed c *, defined as the speed of the biofilm edge along the surface, versus the initial cell cylinder length ℓ 0 for experimental biofilms (A22, magenta; no treatment, yellow; Cefalexin, cyan), agent-based model biofilms (black circles), and continuum model (dashed black curve). Expansion velocities were determined from a linear fit of the basal radius R B of the biofilm versus time, where R B is defined at each time point as the radius of a circle with area equal to that of the biofilm base. For experimental biofilms, the boundary was extracted from the normalized fluorescence data (see Methods for details). For each treatment, the vertical error bars show the standard error of the mean of the expansion speed and the horizontal error bars bound the measured initial cell cylinder length ( Supplementary Fig. 1 ). Inset: model cells with lengths and radii corresponding to the averages for different treatments, ( c ) Biofilm aspect ratio H/R B for experimental biofilms grown under different treatments, where the biofilm height is defined as H = 3 V ∕ 2 R B 2 , the height of a semi-ellipsoid with a circular base of radius R B and volume V equal to that of the biofilm. Error bars show the standard error of the mean. Inset: overlay of biofilm outlines from bottom row of panel ( a ). Color designations and treatments same as in panel ( a ).

    Article Snippet: For biofilm clusters grown in the presence of A22 or Cefalexin, cell lengths were manually measured in the bottom cell layers of the biofilms using the Nikon Element software.

    Techniques: Fluorescence

    Biomass within the biofilm grown on non-functionalized (low-density polyethylene (LDPE) and LDPE-glutaraldehyde (GA)) and functionalized polyethylene surfaces (LDPE-α-chymotrypsin (α-CT)) by plate count viability assay. Data represent the mean ± standard deviation of four independent measurements. Letters a, b and c indicate significant differences (Tukey’s honest significant different (HSD) test, p ≤ 0.05) between the means of different surfaces.

    Journal: International Journal of Molecular Sciences

    Article Title: α-Chymotrypsin Immobilized on a Low-Density Polyethylene Surface Successfully Weakens Escherichia coli Biofilm Formation

    doi: 10.3390/ijms19124003

    Figure Lengend Snippet: Biomass within the biofilm grown on non-functionalized (low-density polyethylene (LDPE) and LDPE-glutaraldehyde (GA)) and functionalized polyethylene surfaces (LDPE-α-chymotrypsin (α-CT)) by plate count viability assay. Data represent the mean ± standard deviation of four independent measurements. Letters a, b and c indicate significant differences (Tukey’s honest significant different (HSD) test, p ≤ 0.05) between the means of different surfaces.

    Article Snippet: Biofilm samples were visualized using a Nikon Eclipse E800 epifluorescent microscope with excitation at 480 nm and emission at 516 nm for the green channel and excitation at 581 nm and emission at 644 nm for the red channel (Tokyo, Japan).

    Techniques: Viability Assay, Standard Deviation

    Confocal laser scanning microscopy analysis. Representative projection analysis ( A , B ) and three-dimensionally (3D) reconstructed CLSM images ( C , D ) of E. coli biofilm grown on non-functionalized LDPE surface ( A , C ) and LDPE-α-CT ( B , D ) functionalized surface (63×, 0.9 NA water immersion objective). The arrows indicate cells detaching from the biofilm. Live cells were stained green with SYBR Green I (λ ex at 488 nm, λ em at 520 nm), whereas the polysaccharide matrix was stained red with Texas Red-labeled concanavalin A (ConA) (λ ex at 543 nm, λ em at 615 nm). ( E ) Representative 3D reconstructed CLSM images of LDPE and LDPE-α-CT without biofilm and stained with SYBR Green I or Texas Red-labeled ConA showing no detectable fluorescence. Scale bar = 20, 30, or 40 µm.

    Journal: International Journal of Molecular Sciences

    Article Title: α-Chymotrypsin Immobilized on a Low-Density Polyethylene Surface Successfully Weakens Escherichia coli Biofilm Formation

    doi: 10.3390/ijms19124003

    Figure Lengend Snippet: Confocal laser scanning microscopy analysis. Representative projection analysis ( A , B ) and three-dimensionally (3D) reconstructed CLSM images ( C , D ) of E. coli biofilm grown on non-functionalized LDPE surface ( A , C ) and LDPE-α-CT ( B , D ) functionalized surface (63×, 0.9 NA water immersion objective). The arrows indicate cells detaching from the biofilm. Live cells were stained green with SYBR Green I (λ ex at 488 nm, λ em at 520 nm), whereas the polysaccharide matrix was stained red with Texas Red-labeled concanavalin A (ConA) (λ ex at 543 nm, λ em at 615 nm). ( E ) Representative 3D reconstructed CLSM images of LDPE and LDPE-α-CT without biofilm and stained with SYBR Green I or Texas Red-labeled ConA showing no detectable fluorescence. Scale bar = 20, 30, or 40 µm.

    Article Snippet: Biofilm samples were visualized using a Nikon Eclipse E800 epifluorescent microscope with excitation at 480 nm and emission at 516 nm for the green channel and excitation at 581 nm and emission at 644 nm for the red channel (Tokyo, Japan).

    Techniques: Confocal Laser Scanning Microscopy, Staining, SYBR Green Assay, Labeling, Fluorescence

    Epifluorescence microscope analysis. ( A ) Percentage of live and dead cells within the biofilm grown on non-functionalized (LDPE and LDPE-GA) and functionalized polyethylene surfaces (LDPE-α-CT). Data represent the mean ± standard deviation of four independent measurements. Letters a, b and c indicate significant differences (Tukey’s HSD, p ≤ 0.05) between the means of different surfaces. ( B – D ) Representative epifluorescence microscope images of E. coli biofilm stained with a Live/Dead BacLight viability kit and grown on LDPE ( B ), LDPE-GA ( C ), and LDPE-α-CT ( D ) surfaces (60×, 1.0 NA water immersion objective). Green fluorescence corresponds to E. coli live cells (λ ex : 480 nm and λ em : 516 nm) and red fluorescence corresponds to E. coli dead cells (λ ex : 581 nm and λ em : 644 nm). ( E ) Representative epifluorescence microscope image of LDPE, LDPE-GA, and LDPE-α-CT without biofilm and stained with a Live/Dead BacLight viability kit showing no detectable fluorescence. Scale bar = 20 µm.

    Journal: International Journal of Molecular Sciences

    Article Title: α-Chymotrypsin Immobilized on a Low-Density Polyethylene Surface Successfully Weakens Escherichia coli Biofilm Formation

    doi: 10.3390/ijms19124003

    Figure Lengend Snippet: Epifluorescence microscope analysis. ( A ) Percentage of live and dead cells within the biofilm grown on non-functionalized (LDPE and LDPE-GA) and functionalized polyethylene surfaces (LDPE-α-CT). Data represent the mean ± standard deviation of four independent measurements. Letters a, b and c indicate significant differences (Tukey’s HSD, p ≤ 0.05) between the means of different surfaces. ( B – D ) Representative epifluorescence microscope images of E. coli biofilm stained with a Live/Dead BacLight viability kit and grown on LDPE ( B ), LDPE-GA ( C ), and LDPE-α-CT ( D ) surfaces (60×, 1.0 NA water immersion objective). Green fluorescence corresponds to E. coli live cells (λ ex : 480 nm and λ em : 516 nm) and red fluorescence corresponds to E. coli dead cells (λ ex : 581 nm and λ em : 644 nm). ( E ) Representative epifluorescence microscope image of LDPE, LDPE-GA, and LDPE-α-CT without biofilm and stained with a Live/Dead BacLight viability kit showing no detectable fluorescence. Scale bar = 20 µm.

    Article Snippet: Biofilm samples were visualized using a Nikon Eclipse E800 epifluorescent microscope with excitation at 480 nm and emission at 516 nm for the green channel and excitation at 581 nm and emission at 644 nm for the red channel (Tokyo, Japan).

    Techniques: Microscopy, Standard Deviation, Staining, Fluorescence

    Biofilm formation by X. axonopodis pv. citri strains. (A) Green fluorescent protein (GFP)-labeled bacteria were grown on chambered cover slides and visualized under confocal laser scanning microscopy (CLSM) after two (i) and five (ii) days of bacterial growth. For each time period, the upper panels show the biofilms developed at the bottom of the chambered cover slides with a magnification of 400X, and the bottom panels show a 2X zoom of the regions marked in the previous panels. Scale bars, 50 µm. In this experiment Δ lov -p lov corresponds to the Δ lov strain complemented with pBBR-p lov 2 (Δ lov -p lov ’). (B) X. axonopodis pv. citri strains were statically grown on glass tubes for two weeks at 28°C. Biofilms were observed on the air-liquid interface. In each case, bacteria were grown under light and dark conditions.

    Journal: PLoS ONE

    Article Title: A LOV Protein Modulates the Physiological Attributes of Xanthomonas axonopodis pv. citri Relevant for Host Plant Colonization

    doi: 10.1371/journal.pone.0038226

    Figure Lengend Snippet: Biofilm formation by X. axonopodis pv. citri strains. (A) Green fluorescent protein (GFP)-labeled bacteria were grown on chambered cover slides and visualized under confocal laser scanning microscopy (CLSM) after two (i) and five (ii) days of bacterial growth. For each time period, the upper panels show the biofilms developed at the bottom of the chambered cover slides with a magnification of 400X, and the bottom panels show a 2X zoom of the regions marked in the previous panels. Scale bars, 50 µm. In this experiment Δ lov -p lov corresponds to the Δ lov strain complemented with pBBR-p lov 2 (Δ lov -p lov ’). (B) X. axonopodis pv. citri strains were statically grown on glass tubes for two weeks at 28°C. Biofilms were observed on the air-liquid interface. In each case, bacteria were grown under light and dark conditions.

    Article Snippet: Biofilm formation was visualized by confocal laser scanning microscopy (Nikon Eclipse TE-2000-E2) with a motor system and DIC/Nomarski optics and a head scan D Eclipse C1si.

    Techniques: Labeling, Confocal Laser Scanning Microscopy

    Characterization of the phenotype of BCR1 -phosphorylation mutants in biofilm formation in vitro . BCR1 phosphomimetic ( bcr1 T191E / bcr1 Δ, JC1094; bcr1 S556E / bcr1 Δ, JC1092; bcr1EE / bcr1 Δ, JC1180) and phosphodefective ( bcr1 T191A / bcr1 Δ, JC1093; bcr1 S556A / bcr1 Δ, JC1088; bcr1AA / bcr1 Δ, JC1178) mutants were grown in biofilm-inducing conditions for 40 hours using microfermentors. BCR1/bcr1 Δ (WT, JC1089) and bcr1 Δ (JC1081) strains were used as controls. (A) Quantification of adherent cells. (B) Biofilm dry mass determination. (C) Biofilms grown using silicon squares models were stained with calcofluor white for CSLM visualization.

    Journal: PLoS Pathogens

    Article Title: The NDR/LATS Kinase Cbk1 Controls the Activity of the Transcriptional Regulator Bcr1 during Biofilm Formation in Candida albicans

    doi: 10.1371/journal.ppat.1002683

    Figure Lengend Snippet: Characterization of the phenotype of BCR1 -phosphorylation mutants in biofilm formation in vitro . BCR1 phosphomimetic ( bcr1 T191E / bcr1 Δ, JC1094; bcr1 S556E / bcr1 Δ, JC1092; bcr1EE / bcr1 Δ, JC1180) and phosphodefective ( bcr1 T191A / bcr1 Δ, JC1093; bcr1 S556A / bcr1 Δ, JC1088; bcr1AA / bcr1 Δ, JC1178) mutants were grown in biofilm-inducing conditions for 40 hours using microfermentors. BCR1/bcr1 Δ (WT, JC1089) and bcr1 Δ (JC1081) strains were used as controls. (A) Quantification of adherent cells. (B) Biofilm dry mass determination. (C) Biofilms grown using silicon squares models were stained with calcofluor white for CSLM visualization.

    Article Snippet: Microscopy and image analysis Biofilm development in microfermentors was recorded with a Nikon Coolpix digital camera.

    Techniques: In Vitro, Staining

    Characterization of the in vitro biofilm formation phenotype of the cbk1 Δ bcr1EE double mutant. (A) Quantification of adherence ability to plastic slides of wild-type (JC1089), cbk1 Δ BCR1 (JC1082), cbk1 Δ bcr1AA (JC1096) and cbk1 Δ bcr1EE (JC1099) cells. (B) The same strains were incubated during 40 hours in microfermentors and biofilm dry mass was determined. (C) CSLM images of biofilms induced using silicone squares after calcofluor white staining. (D) Quantitative RT-PCR measurements of ALS3 and ALS1 transcription levels in the wild-type, cbk1 Δ BCR1 and cbk1 Δ bcr1EE strains normalized using ADE2 .

    Journal: PLoS Pathogens

    Article Title: The NDR/LATS Kinase Cbk1 Controls the Activity of the Transcriptional Regulator Bcr1 during Biofilm Formation in Candida albicans

    doi: 10.1371/journal.ppat.1002683

    Figure Lengend Snippet: Characterization of the in vitro biofilm formation phenotype of the cbk1 Δ bcr1EE double mutant. (A) Quantification of adherence ability to plastic slides of wild-type (JC1089), cbk1 Δ BCR1 (JC1082), cbk1 Δ bcr1AA (JC1096) and cbk1 Δ bcr1EE (JC1099) cells. (B) The same strains were incubated during 40 hours in microfermentors and biofilm dry mass was determined. (C) CSLM images of biofilms induced using silicone squares after calcofluor white staining. (D) Quantitative RT-PCR measurements of ALS3 and ALS1 transcription levels in the wild-type, cbk1 Δ BCR1 and cbk1 Δ bcr1EE strains normalized using ADE2 .

    Article Snippet: Microscopy and image analysis Biofilm development in microfermentors was recorded with a Nikon Coolpix digital camera.

    Techniques: In Vitro, Mutagenesis, Incubation, Staining, Quantitative RT-PCR

    The RAM pathway is required for biofilm formation in vitro . (A) The wild-type (CEC369) and bcr1 Δ (JC1081) reference strains, cbk1 Δ (JC1080), mob2 Δ (JC525), tao3 Δ (JC848), kic1 Δ (JC798) and ace2 Δ (JC343) mutants were incubated during 40 hours in biofilm-inducing conditions on Thermanox plastic slides in microfermentors. (B) Determination of biofilm dry mass collected from microfermentors shown in (A). Average results of two independent experiments done in duplicate are shown. Error bars represent the standard deviation of the data throughout the paper, unless otherwise indicated. (C) Adherence to plastic slides of WT, bcr1Δ , RAM mutants and ace2 Δ, as detailed in Materials and Methods . The results shown are the mean of 3 independent experiments counting 30 fields per strain in each one. (D) Biofilms induced on silicon squares for 60 hours were stained with calcofluor white and concanavaline A for CSLM visualization.

    Journal: PLoS Pathogens

    Article Title: The NDR/LATS Kinase Cbk1 Controls the Activity of the Transcriptional Regulator Bcr1 during Biofilm Formation in Candida albicans

    doi: 10.1371/journal.ppat.1002683

    Figure Lengend Snippet: The RAM pathway is required for biofilm formation in vitro . (A) The wild-type (CEC369) and bcr1 Δ (JC1081) reference strains, cbk1 Δ (JC1080), mob2 Δ (JC525), tao3 Δ (JC848), kic1 Δ (JC798) and ace2 Δ (JC343) mutants were incubated during 40 hours in biofilm-inducing conditions on Thermanox plastic slides in microfermentors. (B) Determination of biofilm dry mass collected from microfermentors shown in (A). Average results of two independent experiments done in duplicate are shown. Error bars represent the standard deviation of the data throughout the paper, unless otherwise indicated. (C) Adherence to plastic slides of WT, bcr1Δ , RAM mutants and ace2 Δ, as detailed in Materials and Methods . The results shown are the mean of 3 independent experiments counting 30 fields per strain in each one. (D) Biofilms induced on silicon squares for 60 hours were stained with calcofluor white and concanavaline A for CSLM visualization.

    Article Snippet: Microscopy and image analysis Biofilm development in microfermentors was recorded with a Nikon Coolpix digital camera.

    Techniques: In Vitro, Incubation, Standard Deviation, Staining