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Millipore biofilms
Effect of NAC on S . maltophilia and Bcc <t>biofilm</t> formation (72-hours growth). Data from at least two independent experiments, with at least four replicates per condition per experiment. Median values with standard deviation are plotted. The x axis is set at the limit of detection (i.e., 1.3 log CFU/peg).
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Images

1) Product Images from "In vitro activity of N-acetylcysteine against Stenotrophomonas maltophilia and Burkholderia cepacia complex grown in planktonic phase and biofilm"

Article Title: In vitro activity of N-acetylcysteine against Stenotrophomonas maltophilia and Burkholderia cepacia complex grown in planktonic phase and biofilm

Journal: PLoS ONE

doi: 10.1371/journal.pone.0203941

Effect of NAC on S . maltophilia and Bcc biofilm formation (72-hours growth). Data from at least two independent experiments, with at least four replicates per condition per experiment. Median values with standard deviation are plotted. The x axis is set at the limit of detection (i.e., 1.3 log CFU/peg).
Figure Legend Snippet: Effect of NAC on S . maltophilia and Bcc biofilm formation (72-hours growth). Data from at least two independent experiments, with at least four replicates per condition per experiment. Median values with standard deviation are plotted. The x axis is set at the limit of detection (i.e., 1.3 log CFU/peg).

Techniques Used: Standard Deviation

Time-kill curves of NAC for 2-day-old biofilms of S . maltophilia and Bcc. Data from at least two independent experiments, with at least four replicates per condition per experiment. Median values with standard deviation are plotted. The x axis is set at the limit of detection (i.e., 1.3 log CFU/peg).
Figure Legend Snippet: Time-kill curves of NAC for 2-day-old biofilms of S . maltophilia and Bcc. Data from at least two independent experiments, with at least four replicates per condition per experiment. Median values with standard deviation are plotted. The x axis is set at the limit of detection (i.e., 1.3 log CFU/peg).

Techniques Used: Standard Deviation

2) Product Images from "Anti-microfouling Activity of Glycomyces sediminimaris UTMC 2460 on Dominant Fouling Bacteria of Iran Marine Habitats"

Article Title: Anti-microfouling Activity of Glycomyces sediminimaris UTMC 2460 on Dominant Fouling Bacteria of Iran Marine Habitats

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2018.03148

The effect of extract on the viability of Kocuria sp. UTMC 2449 and Mesorhizobium sp. UTMC 2518 cells in biofilm state. The lethal activity of the Glycomyces sp. UTMC 2460 metabolite at the lowest biofilm inhibition concentration (100 μg ml -1 ) was resembeled to ciprofloxacin as the control compound with bactericidal activity.
Figure Legend Snippet: The effect of extract on the viability of Kocuria sp. UTMC 2449 and Mesorhizobium sp. UTMC 2518 cells in biofilm state. The lethal activity of the Glycomyces sp. UTMC 2460 metabolite at the lowest biofilm inhibition concentration (100 μg ml -1 ) was resembeled to ciprofloxacin as the control compound with bactericidal activity.

Techniques Used: Activity Assay, Inhibition, Concentration Assay

The biofilm formation of dominant isolated fouling bacteria from immersed artificial platforms in Oman sea detected by crystal violet assay.
Figure Legend Snippet: The biofilm formation of dominant isolated fouling bacteria from immersed artificial platforms in Oman sea detected by crystal violet assay.

Techniques Used: Isolation, Crystal Violet Assay

3) Product Images from "The MNN2 Gene Knockout Modulates the Antifungal Resistance of Biofilms of Candida glabrata"

Article Title: The MNN2 Gene Knockout Modulates the Antifungal Resistance of Biofilms of Candida glabrata

Journal: Biomolecules

doi: 10.3390/biom8040130

Confocal laser scanning microscopy image a 48-h-biofilm of Candida glabrata ATCC2001, C. glabrata ΔHT6 and C. glabrata Δ mnn2 . The biofilm images were acquired using a confocal scanning laser microscope (Olympus BX61, Model FluoView 1000). Filters: DAPI (100 mg/L emissions filters BA 430–470) and Concanavalin A, Alexa Fluor 488 conjugate (50 mg/L emissions filters BA 505–605). Images were acquired with the program FV10-ASW 4.2 (Olympus) using a magnification of 100×. Measure bar: 10 µm.
Figure Legend Snippet: Confocal laser scanning microscopy image a 48-h-biofilm of Candida glabrata ATCC2001, C. glabrata ΔHT6 and C. glabrata Δ mnn2 . The biofilm images were acquired using a confocal scanning laser microscope (Olympus BX61, Model FluoView 1000). Filters: DAPI (100 mg/L emissions filters BA 430–470) and Concanavalin A, Alexa Fluor 488 conjugate (50 mg/L emissions filters BA 505–605). Images were acquired with the program FV10-ASW 4.2 (Olympus) using a magnification of 100×. Measure bar: 10 µm.

Techniques Used: Confocal Laser Scanning Microscopy, Microscopy

Alcian Blue binding assay. Data represent the mean amount of dye bound per biofilm cell of C. glabrata ATCC2001, C. glabrata ΔHT6, and C. glabrata Δ mnn2 (undetected). (Cg— C. glabrata . * p
Figure Legend Snippet: Alcian Blue binding assay. Data represent the mean amount of dye bound per biofilm cell of C. glabrata ATCC2001, C. glabrata ΔHT6, and C. glabrata Δ mnn2 (undetected). (Cg— C. glabrata . * p

Techniques Used: Binding Assay

Crystal Violet in a 48-h-biofilm of C. glabrata ATCC2001, C. glabrata ΔHT6, and C. glabrata Δ mnn2 with and without antifungal agents. The quantification of the biomass is presented by abs/cm 2 . (* p
Figure Legend Snippet: Crystal Violet in a 48-h-biofilm of C. glabrata ATCC2001, C. glabrata ΔHT6, and C. glabrata Δ mnn2 with and without antifungal agents. The quantification of the biomass is presented by abs/cm 2 . (* p

Techniques Used:

4) Product Images from "Bone Environment Influences Irreversible Adhesion of a Methicillin-Susceptible Staphylococcus aureus Strain"

Article Title: Bone Environment Influences Irreversible Adhesion of a Methicillin-Susceptible Staphylococcus aureus Strain

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2018.02865

Hypoxia increased S. aureus biomass biofilm. Planktonic growth normalized on control (A) and biofilm biomass quantified by crystal violet staining (B) under aerobic and anaerobic conditions. ( n = 9). Scanning Electronic Microscopy (SEM) (C) . Main panels: scale bar = 1 μm and insert panels showing the homogeneity on a wide field: scale bar = 10 μm.
Figure Legend Snippet: Hypoxia increased S. aureus biomass biofilm. Planktonic growth normalized on control (A) and biofilm biomass quantified by crystal violet staining (B) under aerobic and anaerobic conditions. ( n = 9). Scanning Electronic Microscopy (SEM) (C) . Main panels: scale bar = 1 μm and insert panels showing the homogeneity on a wide field: scale bar = 10 μm.

Techniques Used: Staining, Microscopy

Synthetic peptides (1018, 1002, 3002, and DJK-5, 5 μg/ml) inhibited osteoblast products effect on S. aureus biofilm formation. Fold-increase of biofilm biomass quantified by crystal violet staining (A,C) and fold-increase of live adhered cells (B,D) under normal conditions of osteoblast-like culture (A,B) and TNF-α stimulated osteoblast-like culture (C,D) . SN cell., increase of biofilm formation in contact of osteoblast supernatants. SN cell. TNF-α, increase of biofilm formation in contact of osteoblast supernatants culture after TNF-α exposition. ( n = 9). ∗ Statistically significantly different from biofilm formed in the presence of SN cell or SN cell. TNF-α ( p
Figure Legend Snippet: Synthetic peptides (1018, 1002, 3002, and DJK-5, 5 μg/ml) inhibited osteoblast products effect on S. aureus biofilm formation. Fold-increase of biofilm biomass quantified by crystal violet staining (A,C) and fold-increase of live adhered cells (B,D) under normal conditions of osteoblast-like culture (A,B) and TNF-α stimulated osteoblast-like culture (C,D) . SN cell., increase of biofilm formation in contact of osteoblast supernatants. SN cell. TNF-α, increase of biofilm formation in contact of osteoblast supernatants culture after TNF-α exposition. ( n = 9). ∗ Statistically significantly different from biofilm formed in the presence of SN cell or SN cell. TNF-α ( p

Techniques Used: Staining

Impact of Mg 2+ on S. aureus biofilm formation. Planktonic growth normalized to control (A) ; biofilm biomass quantified by crystal violet staining (B) and fold-increase of live adhered cells (C) . Mg 2+ × 2 (control) and ×20 serological concentration (=1 mM). ( n = 9). ∗ Statistically significantly different from control ( p
Figure Legend Snippet: Impact of Mg 2+ on S. aureus biofilm formation. Planktonic growth normalized to control (A) ; biofilm biomass quantified by crystal violet staining (B) and fold-increase of live adhered cells (C) . Mg 2+ × 2 (control) and ×20 serological concentration (=1 mM). ( n = 9). ∗ Statistically significantly different from control ( p

Techniques Used: Staining, Concentration Assay

Diluted supernatants of osteoblast culture influenced S. aureus biofilm formation. Planktonic growth normalized on control (A) biomass biofilm quantified by crystal violet staining (B) and fold-increase of live adhered cells (C) . Control media = 50% DMEM + 10% FCS and 50% of minimal medium; SN 50 = culture with 50% of osteoblast culture supernatants and 50% of minimal media. ( n = 9).
Figure Legend Snippet: Diluted supernatants of osteoblast culture influenced S. aureus biofilm formation. Planktonic growth normalized on control (A) biomass biofilm quantified by crystal violet staining (B) and fold-increase of live adhered cells (C) . Control media = 50% DMEM + 10% FCS and 50% of minimal medium; SN 50 = culture with 50% of osteoblast culture supernatants and 50% of minimal media. ( n = 9).

Techniques Used: Staining

Glucose and amino acid starvation increased S. aureus biofilm formation when normalized on planktonic growth. Planktonic growth normalized on control (A) fold-increase of biofilm biomass quantified by crystal violet staining (B) and fold-increase of live adhered cells (C) , fold-increase of biomass fraction on planktonic growth (D) and fold-increase of adherent cells fraction on planktonic growth (E) . Fluorescence microscopy (F) with live (green color)/dead (red color) staining. Scale bar = 40 μm. no iron, without iron; no CAA, without casamino acids; no Glu, without glucose. ( n = 9). ∗ Statistically significantly different from control ( p
Figure Legend Snippet: Glucose and amino acid starvation increased S. aureus biofilm formation when normalized on planktonic growth. Planktonic growth normalized on control (A) fold-increase of biofilm biomass quantified by crystal violet staining (B) and fold-increase of live adhered cells (C) , fold-increase of biomass fraction on planktonic growth (D) and fold-increase of adherent cells fraction on planktonic growth (E) . Fluorescence microscopy (F) with live (green color)/dead (red color) staining. Scale bar = 40 μm. no iron, without iron; no CAA, without casamino acids; no Glu, without glucose. ( n = 9). ∗ Statistically significantly different from control ( p

Techniques Used: Staining, Fluorescence, Microscopy, Cellular Antioxidant Activity Assay

Diluted supernatants of osteoblast culture stimulated with TNF-α influenced S. aureus biofilm formation. Planktonic growth normalized on control (A) fold-increase of biomass biofilm quantified by crystal violet staining (B) and fold-increase of live adhered cells (C) . Control medium + TNF-α = 50% DMEM, 10% FCS, 20 ng/ml TNF-α, 50% minimal medium; SN 50 + TNF-α = culture with 50% of osteoblast culture supernatants exposed to 20 ng/ml of TNF-α and 50% of minimal media. ( n = 9).
Figure Legend Snippet: Diluted supernatants of osteoblast culture stimulated with TNF-α influenced S. aureus biofilm formation. Planktonic growth normalized on control (A) fold-increase of biomass biofilm quantified by crystal violet staining (B) and fold-increase of live adhered cells (C) . Control medium + TNF-α = 50% DMEM, 10% FCS, 20 ng/ml TNF-α, 50% minimal medium; SN 50 + TNF-α = culture with 50% of osteoblast culture supernatants exposed to 20 ng/ml of TNF-α and 50% of minimal media. ( n = 9).

Techniques Used: Staining

Impact of Ca 2+ on S. aureus biofilm formation. Planktonic growth normalized on control (A) and fold-increase of live adhered cells (B) . Ca 2+ × 1 and ×2 serological concentration (=1.2 mM). ( n = 9). ∗ Statistically significantly different from control ( p
Figure Legend Snippet: Impact of Ca 2+ on S. aureus biofilm formation. Planktonic growth normalized on control (A) and fold-increase of live adhered cells (B) . Ca 2+ × 1 and ×2 serological concentration (=1.2 mM). ( n = 9). ∗ Statistically significantly different from control ( p

Techniques Used: Concentration Assay

Dynamic model confirmed the impact of osteoblast culture supernatants on S. aureus biofilm formation. Impact of osteoblast culture on biofilm was tested into the flow-through medium of the flow cell apparatus for 24 h. Each panel shows 3D reconstruction by Imaris software after acquisition in confocal microscopy with live (green color)/dead (red color) staining. Bottom panel represents the quantitative data calculated Imaris software based on acquired images. Control media, 50% DMEM + 10% FCS and 50% of minimal media; SN 50, culture with 50% of osteoblast culture supernatants and 50% of minimal media; Control media + TNF-α = 50% DMEM, 10% FCS, 20 ng/ml TNF-α, 50% minimal media; SN 50+TNF-α, culture with 50% of osteoblast culture supernatants exposed to 20 ng/ml of TNF-α and 50% of minimal media.
Figure Legend Snippet: Dynamic model confirmed the impact of osteoblast culture supernatants on S. aureus biofilm formation. Impact of osteoblast culture on biofilm was tested into the flow-through medium of the flow cell apparatus for 24 h. Each panel shows 3D reconstruction by Imaris software after acquisition in confocal microscopy with live (green color)/dead (red color) staining. Bottom panel represents the quantitative data calculated Imaris software based on acquired images. Control media, 50% DMEM + 10% FCS and 50% of minimal media; SN 50, culture with 50% of osteoblast culture supernatants and 50% of minimal media; Control media + TNF-α = 50% DMEM, 10% FCS, 20 ng/ml TNF-α, 50% minimal media; SN 50+TNF-α, culture with 50% of osteoblast culture supernatants exposed to 20 ng/ml of TNF-α and 50% of minimal media.

Techniques Used: Flow Cytometry, Software, Confocal Microscopy, Staining

5) Product Images from "Effects of Chlorhexidine-Encapsulated Mesoporous Silica Nanoparticles on the Anti-Biofilm and Mechanical Properties of Glass Ionomer Cement"

Article Title: Effects of Chlorhexidine-Encapsulated Mesoporous Silica Nanoparticles on the Anti-Biofilm and Mechanical Properties of Glass Ionomer Cement

Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

doi: 10.3390/molecules22071225

Relative biofilm viability of control, 1%, 5%, and 10% CHX@pMSN groups by MTT assay on ( A ) day 1; and ( B ) day 30. Data are shown as mean ± standard deviation. The groups labeled with the same letters have no significant difference ( p > 0.05).
Figure Legend Snippet: Relative biofilm viability of control, 1%, 5%, and 10% CHX@pMSN groups by MTT assay on ( A ) day 1; and ( B ) day 30. Data are shown as mean ± standard deviation. The groups labeled with the same letters have no significant difference ( p > 0.05).

Techniques Used: MTT Assay, Standard Deviation, Labeling

FESEM and CLSM evaluation of S. mutans biofilms. (Top) FESEM images of ( A ) control; and ( B ) 1% CHX@pMSN groups. (Middle) CLSM 3D overlay projections of ( C ) control; and ( D ) 1% CHX@pMSN groups. (Bottom) Corresponding distribution of live/dead bacteria biomass at each layer of Z-stack from ( E ) control; and ( F ) 1% CHX@pMSN groups.
Figure Legend Snippet: FESEM and CLSM evaluation of S. mutans biofilms. (Top) FESEM images of ( A ) control; and ( B ) 1% CHX@pMSN groups. (Middle) CLSM 3D overlay projections of ( C ) control; and ( D ) 1% CHX@pMSN groups. (Bottom) Corresponding distribution of live/dead bacteria biomass at each layer of Z-stack from ( E ) control; and ( F ) 1% CHX@pMSN groups.

Techniques Used: Confocal Laser Scanning Microscopy

6) Product Images from "Bundle-Forming Pili and EspA Are Involved in Biofilm Formation by Enteropathogenic Escherichia coli"

Article Title: Bundle-Forming Pili and EspA Are Involved in Biofilm Formation by Enteropathogenic Escherichia coli

Journal: Journal of Bacteriology

doi: 10.1128/JB.00177-06

Proposed model of biofilm formation by EPEC.
Figure Legend Snippet: Proposed model of biofilm formation by EPEC.

Techniques Used:

Transcription expression of EPEC virulence genes ( LEE1 , LEE2 , LEE3 , LEE4 , LEE5 , per , bfp , fliA , fliC , and flhDC ) during biofilm formation on plastic in DMEM using transcription fusions with a reporter gfp gene (plasmid pFPV25). We used as a negative control the bla :: gfp (β-lactamase promoter) fusion, which is constitutive. Transcription of all promoters was normalized by the number of bacterial cells and subtracted from basal gfp expression levels expressed by the promoterless gfp vector (pFPV25).
Figure Legend Snippet: Transcription expression of EPEC virulence genes ( LEE1 , LEE2 , LEE3 , LEE4 , LEE5 , per , bfp , fliA , fliC , and flhDC ) during biofilm formation on plastic in DMEM using transcription fusions with a reporter gfp gene (plasmid pFPV25). We used as a negative control the bla :: gfp (β-lactamase promoter) fusion, which is constitutive. Transcription of all promoters was normalized by the number of bacterial cells and subtracted from basal gfp expression levels expressed by the promoterless gfp vector (pFPV25).

Techniques Used: Expressing, Plasmid Preparation, Negative Control

Measurement of biofilm biomass during different incubation time points (3, 6, 9, 12, and 24 h) by WT EPEC (filled diamonds) and isogenic mutants in espA (filled squares), bfp (open triangles), qseA (×'s), fimA (filled circles), and flu (open circles) in DMEM on an abiotic surface under static conditions. Biomass is expressed as CFU/cm 2 . Error bars indicate standard errors of the means.
Figure Legend Snippet: Measurement of biofilm biomass during different incubation time points (3, 6, 9, 12, and 24 h) by WT EPEC (filled diamonds) and isogenic mutants in espA (filled squares), bfp (open triangles), qseA (×'s), fimA (filled circles), and flu (open circles) in DMEM on an abiotic surface under static conditions. Biomass is expressed as CFU/cm 2 . Error bars indicate standard errors of the means.

Techniques Used: Incubation

Light microscopy of biofilm formation by WT EPEC and isogenic mutants (of espA , bfpA , and qseA ) during different incubation time points (6, 12, and 24 h) in DMEM on an abiotic surface under static conditions. Magnification, ×100.
Figure Legend Snippet: Light microscopy of biofilm formation by WT EPEC and isogenic mutants (of espA , bfpA , and qseA ) during different incubation time points (6, 12, and 24 h) in DMEM on an abiotic surface under static conditions. Magnification, ×100.

Techniques Used: Light Microscopy, Incubation

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 "The Antistaphylococcal Activity of Citropin 1.1 and Temporin A against Planktonic Cells and Biofilms Formed by Isolates from Patients with Atopic Dermatitis: An Assessment of Their Potential to Induce Microbial Resistance Compared to Conventional Antimicrobials"

Article Title: The Antistaphylococcal Activity of Citropin 1.1 and Temporin A against Planktonic Cells and Biofilms Formed by Isolates from Patients with Atopic Dermatitis: An Assessment of Their Potential to Induce Microbial Resistance Compared to Conventional Antimicrobials

Journal: Pharmaceuticals

doi: 10.3390/ph9020030

Antistaphylococcal activity of tested compounds towards biofilms formed by tested SA strains (mg/L); *: compound not active at the tested concentrations.
Figure Legend Snippet: Antistaphylococcal activity of tested compounds towards biofilms formed by tested SA strains (mg/L); *: compound not active at the tested concentrations.

Techniques Used: Activity Assay

9) Product Images from "Removing Biofilms from Microstructured Titanium Ex Vivo: A Novel Approach Using Atmospheric Plasma Technology"

Article Title: Removing Biofilms from Microstructured Titanium Ex Vivo: A Novel Approach Using Atmospheric Plasma Technology

Journal: PLoS ONE

doi: 10.1371/journal.pone.0025893

SEM micrographs, fluorescence microscopy images and photographs of contact areas on Rodac plates of 24-h ( Figure 3A ) and 72-h ( Figure 3B ) in situ biofilms. Biofilms were pre-treated with plasma ((either 3 W (a–c) or 5 W (d–f)), air/water sprayed, and treated again with plasma (experimental treatment sequence III). No biofilm remnants could be detected on the microstructured titanium surfaces after this sequential treatment. (Magnification: SEM a, d: ×10,000; FM b, e: ×5). No microbial growth is detectable on Rodac plates after plasma treatment (c, f).
Figure Legend Snippet: SEM micrographs, fluorescence microscopy images and photographs of contact areas on Rodac plates of 24-h ( Figure 3A ) and 72-h ( Figure 3B ) in situ biofilms. Biofilms were pre-treated with plasma ((either 3 W (a–c) or 5 W (d–f)), air/water sprayed, and treated again with plasma (experimental treatment sequence III). No biofilm remnants could be detected on the microstructured titanium surfaces after this sequential treatment. (Magnification: SEM a, d: ×10,000; FM b, e: ×5). No microbial growth is detectable on Rodac plates after plasma treatment (c, f).

Techniques Used: Fluorescence, Microscopy, In Situ, Sequencing

SEM micrographs and photographs of contact areas on Rodac plates (24-h incubation) of untreated (a, b) and plasma treated (c–f) microstructured titanium surfaces without biofilm (experimental treatment sequence IV). Plasma treatment of the titanium surfaces was performed using either a mean plasma jet power of 3 W (c, d) or 5 W (e, f). No surface alterations were detected after plasma treatment of microstructured titanium surfaces. (Magnification: SEM a, c, e: ×10,000). No microbial growth is detectable on Rodac plates after plasma treatment (b, d, f).
Figure Legend Snippet: SEM micrographs and photographs of contact areas on Rodac plates (24-h incubation) of untreated (a, b) and plasma treated (c–f) microstructured titanium surfaces without biofilm (experimental treatment sequence IV). Plasma treatment of the titanium surfaces was performed using either a mean plasma jet power of 3 W (c, d) or 5 W (e, f). No surface alterations were detected after plasma treatment of microstructured titanium surfaces. (Magnification: SEM a, c, e: ×10,000). No microbial growth is detectable on Rodac plates after plasma treatment (b, d, f).

Techniques Used: Incubation, Sequencing

SEM micrographs, fluorescence microscopy images and photographs of contact areas on Rodac plates of untreated (a–c) and plasma treated (d–i) 24-h biofilms ( Figure 1A ) and 72-h biofilms ( Figure 1B ) formed in situ on microstructured titanium surfaces (experimental treatment sequence I). Plasma treatment of the biofilms was performed using either a mean plasma jet power of 3 W (d–f) or 5 W (g–i). The untreated titanium surfaces are covered by a dense bacterial biofilm. After plasma treatment biofilm residues are visible in SEM images on the microstructured titanium surfaces (arrow). Higher red fluorescence of biofilms appeared under FM (Magnification: SEM a, d, g: ×10,000; FM b, e, h: ×100). No microbial growth is detectable on Rodac plates after plasma treatment (c, f, i).
Figure Legend Snippet: SEM micrographs, fluorescence microscopy images and photographs of contact areas on Rodac plates of untreated (a–c) and plasma treated (d–i) 24-h biofilms ( Figure 1A ) and 72-h biofilms ( Figure 1B ) formed in situ on microstructured titanium surfaces (experimental treatment sequence I). Plasma treatment of the biofilms was performed using either a mean plasma jet power of 3 W (d–f) or 5 W (g–i). The untreated titanium surfaces are covered by a dense bacterial biofilm. After plasma treatment biofilm residues are visible in SEM images on the microstructured titanium surfaces (arrow). Higher red fluorescence of biofilms appeared under FM (Magnification: SEM a, d, g: ×10,000; FM b, e, h: ×100). No microbial growth is detectable on Rodac plates after plasma treatment (c, f, i).

Techniques Used: Fluorescence, Microscopy, In Situ, Sequencing

SEM micrographs, fluorescence microscopy images and photographs of contact areas on Rodac plates of 24-h ( Figure 2A ) and 72-h ( Figure 2B ) in situ biofilms treated with air/water spray (a–c) or treated with plasma and subsequent air/water spraying (d–i 2 bar, 5 s, 10 mm distance; treatment subgroups II). Plasma treatment of the biofilms was performed using either a mean plasma jet power of 3 W (d–f) or 5 W (g–i). Air/water spraying does not cause biofilm removal (a–c), however air/water spraying after plasma pre-treatment resulted in nearly biofilm free titanium surfaces (d–i). Only sparse biofilm remnants are visible on the microstructured titanium surfaces (arrows). (Magnification: SEM a, d, g: ×10,000; FM b, e, h: ×5). Microbial growth is detectable on Rodac plates after the first cycle of plasma treatment at 3 W (2B-f).
Figure Legend Snippet: SEM micrographs, fluorescence microscopy images and photographs of contact areas on Rodac plates of 24-h ( Figure 2A ) and 72-h ( Figure 2B ) in situ biofilms treated with air/water spray (a–c) or treated with plasma and subsequent air/water spraying (d–i 2 bar, 5 s, 10 mm distance; treatment subgroups II). Plasma treatment of the biofilms was performed using either a mean plasma jet power of 3 W (d–f) or 5 W (g–i). Air/water spraying does not cause biofilm removal (a–c), however air/water spraying after plasma pre-treatment resulted in nearly biofilm free titanium surfaces (d–i). Only sparse biofilm remnants are visible on the microstructured titanium surfaces (arrows). (Magnification: SEM a, d, g: ×10,000; FM b, e, h: ×5). Microbial growth is detectable on Rodac plates after the first cycle of plasma treatment at 3 W (2B-f).

Techniques Used: Fluorescence, Microscopy, In Situ

10) Product Images from "The Immunomodulatory Drug Glatiramer Acetate is Also an Effective Antimicrobial Agent that Kills Gram-negative Bacteria"

Article Title: The Immunomodulatory Drug Glatiramer Acetate is Also an Effective Antimicrobial Agent that Kills Gram-negative Bacteria

Journal: Scientific Reports

doi: 10.1038/s41598-017-15969-3

Antimicrobial effects of GA and LL-37 against preformed P . aeruginosa biofilms. ( A ) LIVE/DEAD staining of P . aeruginosa (PAO1) biofilms, initially grown for 24 h in IBIDI flow cells and treated for 3 h with 640 µg/ml LL-37 or GA. Living cells are stained with SYTO 9 (green) and dead cells are stained with PI (red). Scale bar 50 µm. ( B ) Quantification of the percentage of live cells in the biofilms after LL-37 or GA treatment or as untreated. Data were from two experiments with each average and standard deviation calculated from 10 images.
Figure Legend Snippet: Antimicrobial effects of GA and LL-37 against preformed P . aeruginosa biofilms. ( A ) LIVE/DEAD staining of P . aeruginosa (PAO1) biofilms, initially grown for 24 h in IBIDI flow cells and treated for 3 h with 640 µg/ml LL-37 or GA. Living cells are stained with SYTO 9 (green) and dead cells are stained with PI (red). Scale bar 50 µm. ( B ) Quantification of the percentage of live cells in the biofilms after LL-37 or GA treatment or as untreated. Data were from two experiments with each average and standard deviation calculated from 10 images.

Techniques Used: Staining, Flow Cytometry, Standard Deviation

11) Product Images from "Evaluation of combined growth media for in vitro cultivation of oropharyngeal biofilms on prosthetic silicone"

Article Title: Evaluation of combined growth media for in vitro cultivation of oropharyngeal biofilms on prosthetic silicone

Journal: Journal of Materials Science. Materials in Medicine

doi: 10.1007/s10856-018-6051-7

Scheme of the applied microtiter biofilm model and quantification of macroscopic size of biofilm deposits using image analysis software. The size of biofilm cover was calculated as percentage of each total platelet surface. (Artwork created with Inkscape 0.91)
Figure Legend Snippet: Scheme of the applied microtiter biofilm model and quantification of macroscopic size of biofilm deposits using image analysis software. The size of biofilm cover was calculated as percentage of each total platelet surface. (Artwork created with Inkscape 0.91)

Techniques Used: Software

Scanning electron micrographs of the resulting biofilm structures assessed on silicone after 22 days in the in vitro biofilm model. Key structures of biofilms on explanted voice prostheses, such as budded yeast, hypheal germination and balanced bacterial proliferation were assessed in all growth media tested, except in the mixture of FBS 30% + YNB 70%
Figure Legend Snippet: Scanning electron micrographs of the resulting biofilm structures assessed on silicone after 22 days in the in vitro biofilm model. Key structures of biofilms on explanted voice prostheses, such as budded yeast, hypheal germination and balanced bacterial proliferation were assessed in all growth media tested, except in the mixture of FBS 30% + YNB 70%

Techniques Used: In Vitro

Overview of growth kinetics of biofilm deposits over 22 days in the tested growth media: yeast peptone dextrose (YPD), fetal bovine serum (FBS) and mixtures with brain heart infusion (BHI), yeast nitrogen base (YNB) and RPMI 1640 (RPMI)
Figure Legend Snippet: Overview of growth kinetics of biofilm deposits over 22 days in the tested growth media: yeast peptone dextrose (YPD), fetal bovine serum (FBS) and mixtures with brain heart infusion (BHI), yeast nitrogen base (YNB) and RPMI 1640 (RPMI)

Techniques Used:

a View of a dysfunctional Provox 2 voice prosthesis at explantation after 96 days in situ. The valve flap and esophageal flange are overgrown by candida based mixed biofilm deposits. Transprosthetic leakage of esophageal contents is caused by impairment of the valve closure due to biofilm deposits and deterioration of the silicone material. b Candida species infiltrate the silicone material of a VP after weeks (cross section of a Provox 2 after 48 weeks in situ, white arrow: site of biofilm infiltration, shaped arrow: surface of the silicone material)
Figure Legend Snippet: a View of a dysfunctional Provox 2 voice prosthesis at explantation after 96 days in situ. The valve flap and esophageal flange are overgrown by candida based mixed biofilm deposits. Transprosthetic leakage of esophageal contents is caused by impairment of the valve closure due to biofilm deposits and deterioration of the silicone material. b Candida species infiltrate the silicone material of a VP after weeks (cross section of a Provox 2 after 48 weeks in situ, white arrow: site of biofilm infiltration, shaped arrow: surface of the silicone material)

Techniques Used: In Situ

12) Product Images from "Development of a flow chamber system for the reproducible in vitro analysis of biofilm formation on implant materials"

Article Title: Development of a flow chamber system for the reproducible in vitro analysis of biofilm formation on implant materials

Journal: PLoS ONE

doi: 10.1371/journal.pone.0172095

3D reconstruction of biofilms in side view. The cells were stained live/dead and analysed by CLSM. Vital cells are depicted in yellow, viable cells in blue. A) S . gordonii , B) S . oralis , C) S . salivarius , D) P . gingivalis , and E) A . actinomycetemcomitans .
Figure Legend Snippet: 3D reconstruction of biofilms in side view. The cells were stained live/dead and analysed by CLSM. Vital cells are depicted in yellow, viable cells in blue. A) S . gordonii , B) S . oralis , C) S . salivarius , D) P . gingivalis , and E) A . actinomycetemcomitans .

Techniques Used: Staining, Confocal Laser Scanning Microscopy

Biofilm heights on titanium substrata in the flow chamber system. In each diagram, the mean biofilm heights for five independent experiments are shown. A = S . gordonii , B = S . oralis , C = S . salivarius , D = P . gingivalis , and E = A . actinomycetemcomitans .
Figure Legend Snippet: Biofilm heights on titanium substrata in the flow chamber system. In each diagram, the mean biofilm heights for five independent experiments are shown. A = S . gordonii , B = S . oralis , C = S . salivarius , D = P . gingivalis , and E = A . actinomycetemcomitans .

Techniques Used: Flow Cytometry

13) Product Images from "Flagellar motility, extracellular proteases and Vibrio cholerae detachment from abiotic and biotic surfaces"

Article Title: Flagellar motility, extracellular proteases and Vibrio cholerae detachment from abiotic and biotic surfaces

Journal: Microbial pathogenesis

doi: 10.1016/j.micpath.2017.10.016

Role of motility in V. cholerae detachments and biofilm dispersal Left and middle panels . Strain C7258 (Wt) and C7258ΔAB (Δ pomAB ) containing plasmid pCMW75 were allowed to form biofilms in LB medium containing IPTG for 8 h at 30°C. At this time point, the preformed biofilms were treated with Q24DA (10 μg/mL) in dimethyl sulfoxide (DMSO) or DMSO (vehicle control) and further incubated up to 64 h. Then, biofilm formation was determined as described in materials and methods. Rightmost panel . Mixtures of strain C7258 (Wt) and C7258ΔABL (Δ pomAB ) at a 1:1 ratio (based on colony forming units) were allowed to form biofilms for different time periods, the biofilms were disrupted with glass beads and the fraction of motile to non-motile cells determined by dilution plating in LB agar containing X-gal. The trendline indicates the decline in motile cells in the aging biofilm. Each bar represents the average of at least three independent experiments. Error bars indicate the standard deviation (STDEV). Symbols: * p
Figure Legend Snippet: Role of motility in V. cholerae detachments and biofilm dispersal Left and middle panels . Strain C7258 (Wt) and C7258ΔAB (Δ pomAB ) containing plasmid pCMW75 were allowed to form biofilms in LB medium containing IPTG for 8 h at 30°C. At this time point, the preformed biofilms were treated with Q24DA (10 μg/mL) in dimethyl sulfoxide (DMSO) or DMSO (vehicle control) and further incubated up to 64 h. Then, biofilm formation was determined as described in materials and methods. Rightmost panel . Mixtures of strain C7258 (Wt) and C7258ΔABL (Δ pomAB ) at a 1:1 ratio (based on colony forming units) were allowed to form biofilms for different time periods, the biofilms were disrupted with glass beads and the fraction of motile to non-motile cells determined by dilution plating in LB agar containing X-gal. The trendline indicates the decline in motile cells in the aging biofilm. Each bar represents the average of at least three independent experiments. Error bars indicate the standard deviation (STDEV). Symbols: * p

Techniques Used: Plasmid Preparation, Incubation, Standard Deviation

Proteolytic activity of V. cholerae biofilms a. Strains C7258 (Wt) and C7258H ( hapA ) were allowed to form biofilms for different time periods. Planktonic cells were discarded and the total proteolytic activity of the intact biofilm determined as described in materials and methods using a fluorescent probe. The biofilm mass was determined by crystal violet staining and the results expressed as fluorescence units (FI) normalized by the biofilm mass (OD 570 ). b . Strains C7258 (Wt) and C7258H ( hapA ) were allowed to form biofilms in borosilicate tubes. Biofilm mass was estimated by crystal violet staining and the proteolytic activity present in the disrupted biofilms was measured using a fluorescent probe as described in materials and methods. Each bar represents the average of at least three independent experiments. Error bars indicate the STDEV. Symbols: ** p
Figure Legend Snippet: Proteolytic activity of V. cholerae biofilms a. Strains C7258 (Wt) and C7258H ( hapA ) were allowed to form biofilms for different time periods. Planktonic cells were discarded and the total proteolytic activity of the intact biofilm determined as described in materials and methods using a fluorescent probe. The biofilm mass was determined by crystal violet staining and the results expressed as fluorescence units (FI) normalized by the biofilm mass (OD 570 ). b . Strains C7258 (Wt) and C7258H ( hapA ) were allowed to form biofilms in borosilicate tubes. Biofilm mass was estimated by crystal violet staining and the proteolytic activity present in the disrupted biofilms was measured using a fluorescent probe as described in materials and methods. Each bar represents the average of at least three independent experiments. Error bars indicate the STDEV. Symbols: ** p

Techniques Used: Activity Assay, Staining, Fluorescence

Role of proteases in the stability of V. cholerae biofilms Strains C7258 (Wt) and C7258H ( hapA ) transformed with plasmid pCMW75 were allowed to form biofilms in the presence of IPTG to enhance the c-di-GMP pool for different time periods in microtiter plates. The amount of crystal violet stained material was measured at OD 570 . The quantity of bacteria attached at each time point was normalized by the amount present at 8 h (relative attachment). The cOmplete protease inhibitor cocktail was added as recommended by the provider to the Wt and hapA strain at 8 h. Each bar represents the average of at least three independent experiments. Error bars indicate the STDEV. Statistical significance was determined using an unpaired T test (* p
Figure Legend Snippet: Role of proteases in the stability of V. cholerae biofilms Strains C7258 (Wt) and C7258H ( hapA ) transformed with plasmid pCMW75 were allowed to form biofilms in the presence of IPTG to enhance the c-di-GMP pool for different time periods in microtiter plates. The amount of crystal violet stained material was measured at OD 570 . The quantity of bacteria attached at each time point was normalized by the amount present at 8 h (relative attachment). The cOmplete protease inhibitor cocktail was added as recommended by the provider to the Wt and hapA strain at 8 h. Each bar represents the average of at least three independent experiments. Error bars indicate the STDEV. Statistical significance was determined using an unpaired T test (* p

Techniques Used: Transformation Assay, Plasmid Preparation, Staining, Protease Inhibitor

Validation of balanced lethal plasmid system to increase c-di-GMP pool late in infection a . Strain C7258ΔT containing plasmid pAJA1thyA-8* (open bar) or pAJA1thyA-8 (filled bar) were allowed to form biofilms in polystyrene and mucin-coated polystyrene microtiter plates as described in materials and methods. b . Strains C7258ΔT, C7258ΔTAB and C7258HΔT containing plasmid pAJA1thyA-8* (open bars) or pAJA1thyA-8 (filled bars) were grown to stationary phase in TSB medium and production of HapA measured as described in methods. Each bar represents the average of three independent cultures and error bars denote the STDEV (** p
Figure Legend Snippet: Validation of balanced lethal plasmid system to increase c-di-GMP pool late in infection a . Strain C7258ΔT containing plasmid pAJA1thyA-8* (open bar) or pAJA1thyA-8 (filled bar) were allowed to form biofilms in polystyrene and mucin-coated polystyrene microtiter plates as described in materials and methods. b . Strains C7258ΔT, C7258ΔTAB and C7258HΔT containing plasmid pAJA1thyA-8* (open bars) or pAJA1thyA-8 (filled bars) were grown to stationary phase in TSB medium and production of HapA measured as described in methods. Each bar represents the average of three independent cultures and error bars denote the STDEV (** p

Techniques Used: Plasmid Preparation, Infection

14) Product Images from "A Novel Antifungal Is Active against Candida albicans Biofilms and Inhibits Mutagenic Acetaldehyde Production In Vitro"

Article Title: A Novel Antifungal Is Active against Candida albicans Biofilms and Inhibits Mutagenic Acetaldehyde Production In Vitro

Journal: PLoS ONE

doi: 10.1371/journal.pone.0097864

Microscopic examination of C. albicans biofilms exposed to HICA. SEM images were taken of biofilms which were grown for 24% HICA for another 24 h at pH 5.2 (A) or pH 7.4 (B). Control images were taken of biofilms grown in RPMI-medium without HICA at pH 5.2 (C) or pH 7.4 (D). Insets show hyphal structures in detail. Scale bars indicate 10 µm in main images and 5 µm in insets.
Figure Legend Snippet: Microscopic examination of C. albicans biofilms exposed to HICA. SEM images were taken of biofilms which were grown for 24% HICA for another 24 h at pH 5.2 (A) or pH 7.4 (B). Control images were taken of biofilms grown in RPMI-medium without HICA at pH 5.2 (C) or pH 7.4 (D). Insets show hyphal structures in detail. Scale bars indicate 10 µm in main images and 5 µm in insets.

Techniques Used:

Changes in metabolic activity and biomass of C. albicans biofilms. Metabolic activities measured by XTT-assay (panel A, B) and dsDNA levels (panel C, D) reflecting the biomass of C. albicans biofilms at pH 5.2 and 7.4. Biofilms were grown for 4, 24 or 48 h in RPMI at pH 7.4 and then exposed to PBS, caspofungin, HICA, leucine, cysteine or ethanol for another 24 h. Values were measured twice in triplicate and expressed as mean (±SEM). Means were compared to the control treatment (RPMI). Statistically significant differences were calculated using a GEE-model and were marked (**p
Figure Legend Snippet: Changes in metabolic activity and biomass of C. albicans biofilms. Metabolic activities measured by XTT-assay (panel A, B) and dsDNA levels (panel C, D) reflecting the biomass of C. albicans biofilms at pH 5.2 and 7.4. Biofilms were grown for 4, 24 or 48 h in RPMI at pH 7.4 and then exposed to PBS, caspofungin, HICA, leucine, cysteine or ethanol for another 24 h. Values were measured twice in triplicate and expressed as mean (±SEM). Means were compared to the control treatment (RPMI). Statistically significant differences were calculated using a GEE-model and were marked (**p

Techniques Used: Activity Assay, XTT Assay

ACH production by C. albicans biofilms. Mean (±SEM) ACH production by C. albicans biofilms. Biofilms at three different stages of growth (4 h, 24 h and 48 h), were incubated for 30 min at 37°C with 100 mM D-glucose (A,B) or 0.05% ethanol (C,D) after 24 h exposure to RPMI, ethanol, caspofungin or HICA at pH 5.2 or 7.4. Values were measured twice in triplicate and means were compared at each time point to the control treatment (RPMI). Statistically significant differences were calculated using a GEE-model and were marked (**p
Figure Legend Snippet: ACH production by C. albicans biofilms. Mean (±SEM) ACH production by C. albicans biofilms. Biofilms at three different stages of growth (4 h, 24 h and 48 h), were incubated for 30 min at 37°C with 100 mM D-glucose (A,B) or 0.05% ethanol (C,D) after 24 h exposure to RPMI, ethanol, caspofungin or HICA at pH 5.2 or 7.4. Values were measured twice in triplicate and means were compared at each time point to the control treatment (RPMI). Statistically significant differences were calculated using a GEE-model and were marked (**p

Techniques Used: Incubation

Relative expression of genes related to ACH metabolism. (A) A schematic model of central carbon metabolism incorporated with genes of interest within ethanol metabolism (abbreviations as follows: G6P = glucose-6-phosphate, KICA = α-ketoisocaproic acid). (B) Heat map panel of gene expression in C. albicans biofilms at three different stages of growth at pH 5.2 and 7.4 after 24 h exposure to PBS, caspofungin, HICA, leucine, cysteine or ethanol. Fold changes are expressed relative to control (RPMI) at the corresponding time point. Black represents no change in expression, green is up-regulation, and red is down-regulation. A brighter color indicates greater degree of change in expression. Relative gene expressions were calculated by the Pfaffl method using the REST 2009 software provided by Qiagen [63] .
Figure Legend Snippet: Relative expression of genes related to ACH metabolism. (A) A schematic model of central carbon metabolism incorporated with genes of interest within ethanol metabolism (abbreviations as follows: G6P = glucose-6-phosphate, KICA = α-ketoisocaproic acid). (B) Heat map panel of gene expression in C. albicans biofilms at three different stages of growth at pH 5.2 and 7.4 after 24 h exposure to PBS, caspofungin, HICA, leucine, cysteine or ethanol. Fold changes are expressed relative to control (RPMI) at the corresponding time point. Black represents no change in expression, green is up-regulation, and red is down-regulation. A brighter color indicates greater degree of change in expression. Relative gene expressions were calculated by the Pfaffl method using the REST 2009 software provided by Qiagen [63] .

Techniques Used: Expressing, Software

15) Product Images from "A Combined Pharmacodynamic Quantitative and Qualitative Model Reveals the Potent Activity of Daptomycin and Delafloxacin against Staphylococcus aureus Biofilms"

Article Title: A Combined Pharmacodynamic Quantitative and Qualitative Model Reveals the Potent Activity of Daptomycin and Delafloxacin against Staphylococcus aureus Biofilms

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.00181-13

Characterization of the biofilm model with MSSA ATCC 25923. (Left) Evolution over time of the crystal violet absorbance (as a marker of biofilm production) and of resorufin fluorescence (as a marker of bacterial viability) for an initial inoculum with
Figure Legend Snippet: Characterization of the biofilm model with MSSA ATCC 25923. (Left) Evolution over time of the crystal violet absorbance (as a marker of biofilm production) and of resorufin fluorescence (as a marker of bacterial viability) for an initial inoculum with

Techniques Used: Marker, Fluorescence

Antibiotic activities against 6-h S. aureus biofilms.
Figure Legend Snippet: Antibiotic activities against 6-h S. aureus biofilms.

Techniques Used:

(Left and middle) Three-dimensional images from confocal laser scanning microscopy of 24-h biofilms of MSSA ATCC 25923 (left) and MRSA ATCC 33591 (middle) under control conditions or after exposure to selected antibiotics at 32 times their MICs for 48
Figure Legend Snippet: (Left and middle) Three-dimensional images from confocal laser scanning microscopy of 24-h biofilms of MSSA ATCC 25923 (left) and MRSA ATCC 33591 (middle) under control conditions or after exposure to selected antibiotics at 32 times their MICs for 48

Techniques Used: Confocal Laser Scanning Microscopy

Concentration-response activities of antibiotics against 6-h biofilms of MSSA ATCC 25923 (left) or MRSA ATCC 33591 (right). The 6-h biofilms were incubated with increasing concentrations of antibiotics (shown on the x axis) for 24 h. The ordinate shows
Figure Legend Snippet: Concentration-response activities of antibiotics against 6-h biofilms of MSSA ATCC 25923 (left) or MRSA ATCC 33591 (right). The 6-h biofilms were incubated with increasing concentrations of antibiotics (shown on the x axis) for 24 h. The ordinate shows

Techniques Used: Concentration Assay, Incubation

Concentration-response activities of antibiotics against 24-h biofilms of MSSA ATCC 25923 (left) or MRSA ATCC 33591 (right). The 24-h biofilms were incubated with increasing concentrations of antibiotics (shown on the x axis) for 48 h. The ordinate shows
Figure Legend Snippet: Concentration-response activities of antibiotics against 24-h biofilms of MSSA ATCC 25923 (left) or MRSA ATCC 33591 (right). The 24-h biofilms were incubated with increasing concentrations of antibiotics (shown on the x axis) for 48 h. The ordinate shows

Techniques Used: Concentration Assay, Incubation

(Left) Three-dimensional images from confocal laser scanning microscopy of 24-h biofilms of MSSA ATCC 25923 (top) or MRSA ATCC 33591 (bottom) under control conditions or after exposure to delafloxacin or daptomycin at 8 and 16 times the respective MIC
Figure Legend Snippet: (Left) Three-dimensional images from confocal laser scanning microscopy of 24-h biofilms of MSSA ATCC 25923 (top) or MRSA ATCC 33591 (bottom) under control conditions or after exposure to delafloxacin or daptomycin at 8 and 16 times the respective MIC

Techniques Used: Confocal Laser Scanning Microscopy

Antibiotic activity against 24-h S. aureus biofilms.
Figure Legend Snippet: Antibiotic activity against 24-h S. aureus biofilms.

Techniques Used: Activity Assay

16) Product Images from "Presence of extracellular DNA in the Candida albicans biofilm matrix and its contribution to biofilms"

Article Title: Presence of extracellular DNA in the Candida albicans biofilm matrix and its contribution to biofilms

Journal: Mycopathologia

doi: 10.1007/s11046-009-9264-y

Example of microscopy evaluation of the effect of DNase treatment on 24 h C.albicans biofilm formation. Microphotographs of cells in wells before (I) and after (II) aspiration of medium and subsequent washings with PBS and crystal violet staining for biofilms treated with 0 mg/ml (a and c) and 0.13 mg/ml of DNase (b and d). The bar in the picture represents 200 μm.
Figure Legend Snippet: Example of microscopy evaluation of the effect of DNase treatment on 24 h C.albicans biofilm formation. Microphotographs of cells in wells before (I) and after (II) aspiration of medium and subsequent washings with PBS and crystal violet staining for biofilms treated with 0 mg/ml (a and c) and 0.13 mg/ml of DNase (b and d). The bar in the picture represents 200 μm.

Techniques Used: Microscopy, Staining

Effect of DNase treatment on C. albicans biofilm formation. DifferentDNase concentrations (0, 0.02, 0.03, 0.06, 0.13, 0.25, 0.50, 1.00 and 2.00 mg/ml) were added to C. albicans cells at different times (0, 1, 2 and 24 h) post-inoculation in the wells of microtiter plates and incubated at 37°C under static conditions. The extent of biofilm formation was estimated by the crystal violet assay. Presented values are mean A 550 ± standard error of mean of four independent experiments with three to eight replicates. Statistically significant differences (compared to biofilms formed in the absence of DNase) are indicated with an asterisk. (*, P
Figure Legend Snippet: Effect of DNase treatment on C. albicans biofilm formation. DifferentDNase concentrations (0, 0.02, 0.03, 0.06, 0.13, 0.25, 0.50, 1.00 and 2.00 mg/ml) were added to C. albicans cells at different times (0, 1, 2 and 24 h) post-inoculation in the wells of microtiter plates and incubated at 37°C under static conditions. The extent of biofilm formation was estimated by the crystal violet assay. Presented values are mean A 550 ± standard error of mean of four independent experiments with three to eight replicates. Statistically significant differences (compared to biofilms formed in the absence of DNase) are indicated with an asterisk. (*, P

Techniques Used: Incubation, Crystal Violet Assay

Effect of addition of exogenous DNA on C. albicans biofilm formation. Different eDNA concentrations (0, 20, 40, 80, 160, 320, 640, 1280 and 2560 ng/ml) were added to C. albicans cells at different times (0, 1, 2 and 24 h) post -inoculation in the wells of microtiter plates and incubated at 37°C under static conditions. The extent of biofilm formation was estimated by the crystal violet assay. Presented values are mean A 550 ± standard error of mean of four independent experiments with three to eight replicates. Statistically significant differences (compared to biofilms formed in the absence of DNA) are indicated with an asterisk. (*, P
Figure Legend Snippet: Effect of addition of exogenous DNA on C. albicans biofilm formation. Different eDNA concentrations (0, 20, 40, 80, 160, 320, 640, 1280 and 2560 ng/ml) were added to C. albicans cells at different times (0, 1, 2 and 24 h) post -inoculation in the wells of microtiter plates and incubated at 37°C under static conditions. The extent of biofilm formation was estimated by the crystal violet assay. Presented values are mean A 550 ± standard error of mean of four independent experiments with three to eight replicates. Statistically significant differences (compared to biofilms formed in the absence of DNA) are indicated with an asterisk. (*, P

Techniques Used: Incubation, Crystal Violet Assay

17) Product Images from "Real-Time Monitoring of nfxB Mutant Occurrence and Dynamics in Pseudomonas aeruginosa Biofilm Exposed to Subinhibitory Concentrations of Ciprofloxacin"

Article Title: Real-Time Monitoring of nfxB Mutant Occurrence and Dynamics in Pseudomonas aeruginosa Biofilm Exposed to Subinhibitory Concentrations of Ciprofloxacin

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.02292-16

Rapid development of nfxB mutants in a 24-h-old PAO1 flow cell biofilm treated with low-dose CIP. The biofilms of PAO1- mCherry -P CD - gfp + were grown in flow cell chambers with a continuous flow of minimal medium for 24 h at 37°C and then grown for an additional 24 h with 0.2 μg/ml CIP (24 h +CIP for 24 h) or without CIP (24 h −CIP for 24 h). Red shows wild-type cells (elongated or filamentous) due to the constitutive expression of mCherry, and green shows nfxB mutants due to the expression of GFP via the P CD - gfp + reporter. z-stacks were generated by using a Zeiss LSM 710 microscope and processed with Imaris 8.2 software (Bitplane). Top-row images show orthogonal 3D views, and middle and bottom (for treated biofilms only) rows show perspective 3D views of biofilms with an overlay of red and green fluorescence. The images shown are representative of results for two independent flow cell channels.
Figure Legend Snippet: Rapid development of nfxB mutants in a 24-h-old PAO1 flow cell biofilm treated with low-dose CIP. The biofilms of PAO1- mCherry -P CD - gfp + were grown in flow cell chambers with a continuous flow of minimal medium for 24 h at 37°C and then grown for an additional 24 h with 0.2 μg/ml CIP (24 h +CIP for 24 h) or without CIP (24 h −CIP for 24 h). Red shows wild-type cells (elongated or filamentous) due to the constitutive expression of mCherry, and green shows nfxB mutants due to the expression of GFP via the P CD - gfp + reporter. z-stacks were generated by using a Zeiss LSM 710 microscope and processed with Imaris 8.2 software (Bitplane). Top-row images show orthogonal 3D views, and middle and bottom (for treated biofilms only) rows show perspective 3D views of biofilms with an overlay of red and green fluorescence. The images shown are representative of results for two independent flow cell channels.

Techniques Used: Flow Cytometry, Expressing, Generated, Microscopy, Software, Fluorescence

Development of nfxB mutants in a 72-h-old PAO1 flow cell biofilm during treatment with low-dose CIP. The biofilms of PAO1- mCherry -P CD - gfp + were grown in three independent channels of flow cell chambers with a continuous flow of minimal medium for 72 h and then treated with 0.2 μg/ml CIP for a total of 96 h. Imaging by CLSM was done every 24 h. At least 3 images were taken per channel at every time point. Red represents wild-type cells due to the constitutive expression of mCherry, and green shows nfxB mutants due to the expression of GFP+ via the P CD - gfp + reporter. z-stacks were generated by using a Zeiss LSM 710 microscope and processed with Imaris 8.2 software (Bitplane). The images show orthogonal 3D biofilm views (left) or a perspective view (right) with an overlay of red and green channel fluorescence.
Figure Legend Snippet: Development of nfxB mutants in a 72-h-old PAO1 flow cell biofilm during treatment with low-dose CIP. The biofilms of PAO1- mCherry -P CD - gfp + were grown in three independent channels of flow cell chambers with a continuous flow of minimal medium for 72 h and then treated with 0.2 μg/ml CIP for a total of 96 h. Imaging by CLSM was done every 24 h. At least 3 images were taken per channel at every time point. Red represents wild-type cells due to the constitutive expression of mCherry, and green shows nfxB mutants due to the expression of GFP+ via the P CD - gfp + reporter. z-stacks were generated by using a Zeiss LSM 710 microscope and processed with Imaris 8.2 software (Bitplane). The images show orthogonal 3D biofilm views (left) or a perspective view (right) with an overlay of red and green channel fluorescence.

Techniques Used: Flow Cytometry, Imaging, Confocal Laser Scanning Microscopy, Expressing, Generated, Microscopy, Software, Fluorescence

18) Product Images from "Structural dynamics of RbmA governs plasticity of Vibrio cholerae biofilms"

Article Title: Structural dynamics of RbmA governs plasticity of Vibrio cholerae biofilms

Journal: eLife

doi: 10.7554/eLife.26163

Models of various RbmA variants and their involvement in biofilm formation. ( A ) Cartoon models representing the various RbmA oligomerization and switch states. The VPS binding is indicated in blue on the FnIII-2 domain (white); dashed line indicates flexible or unfolded protein. The binary switch is represented exclusively in the open state (D-loop conformation) in the D97A/D97K mutants, and at partial occupancy in the WT protein, while the R234A mutant depicts the binary switch in its closed state (O-loop conformation). Dimeric forms of RbmA (WT, R234A) are drawn in their elongated form, as supported by small angle x-ray scattering studies in solution ( Giglio et al., 2013 ). ( B ) Cell organization and RbmA localization in the biofilms of rugose, Δ rbmA and various rbmA mutants. RbmA, represented in purple, is distributed between the surface and the center of the biofilms in rugose and D97 mutants; while in R234A mutant, RbmA localizes at the center of the biofilm with decrease abundance towards the surface. VPS represented in blue and enhanced VPS localization around the biofilm cluster is represented as a thick blue line.
Figure Legend Snippet: Models of various RbmA variants and their involvement in biofilm formation. ( A ) Cartoon models representing the various RbmA oligomerization and switch states. The VPS binding is indicated in blue on the FnIII-2 domain (white); dashed line indicates flexible or unfolded protein. The binary switch is represented exclusively in the open state (D-loop conformation) in the D97A/D97K mutants, and at partial occupancy in the WT protein, while the R234A mutant depicts the binary switch in its closed state (O-loop conformation). Dimeric forms of RbmA (WT, R234A) are drawn in their elongated form, as supported by small angle x-ray scattering studies in solution ( Giglio et al., 2013 ). ( B ) Cell organization and RbmA localization in the biofilms of rugose, Δ rbmA and various rbmA mutants. RbmA, represented in purple, is distributed between the surface and the center of the biofilms in rugose and D97 mutants; while in R234A mutant, RbmA localizes at the center of the biofilm with decrease abundance towards the surface. VPS represented in blue and enhanced VPS localization around the biofilm cluster is represented as a thick blue line.

Techniques Used: Binding Assay, Mutagenesis

Nearest-neighbor distance, vertical orientation, radial orientation and volume of individual cell in the biofilms formed by rugose parental strain and various mutants, analyzed at single-cell resolution. In each row, the same biofilm of a particular strain, indicated on the left, is shown as an x-z -projection (see Figure 4B for a definition of the coordinate system). In each column of the figure, each cell inside a particular biofilm is colored according to either the nearest neighbor distance of the cell, the vertical orientation of the cell, the radial orientation of the cell, and the volume of each cell. The nearest neighbor distance was calculated from cell centroid to cell centroid distances. The vertical orientation of each cell was calculated as the angle of the major axis of the cell with the vertical z -direction. The radial orientation of each cell was calculated as the angle of the cell with the radial vector in spherical coordinates, with an origin at the center of mass in the x-y -plane of the biofilm. Scale bar, 20 μm.
Figure Legend Snippet: Nearest-neighbor distance, vertical orientation, radial orientation and volume of individual cell in the biofilms formed by rugose parental strain and various mutants, analyzed at single-cell resolution. In each row, the same biofilm of a particular strain, indicated on the left, is shown as an x-z -projection (see Figure 4B for a definition of the coordinate system). In each column of the figure, each cell inside a particular biofilm is colored according to either the nearest neighbor distance of the cell, the vertical orientation of the cell, the radial orientation of the cell, and the volume of each cell. The nearest neighbor distance was calculated from cell centroid to cell centroid distances. The vertical orientation of each cell was calculated as the angle of the major axis of the cell with the vertical z -direction. The radial orientation of each cell was calculated as the angle of the cell with the radial vector in spherical coordinates, with an origin at the center of mass in the x-y -plane of the biofilm. Scale bar, 20 μm.

Techniques Used: Plasmid Preparation

Quantitative analysis of ( A ) biofilm biomass, ( B ) average biofilm thickness, ( C ) maximum biofilm thickness, and ( D ) colony counts (CFU/mL) in the effluent of the biofilms formed by the rugose strain, Δ rbmA and various mutant strains at 24 hr post-inoculation as determined by COMSTAT and dilution plating analysis. ***p
Figure Legend Snippet: Quantitative analysis of ( A ) biofilm biomass, ( B ) average biofilm thickness, ( C ) maximum biofilm thickness, and ( D ) colony counts (CFU/mL) in the effluent of the biofilms formed by the rugose strain, Δ rbmA and various mutant strains at 24 hr post-inoculation as determined by COMSTAT and dilution plating analysis. ***p

Techniques Used: Mutagenesis

19) Product Images from "Transcriptome Assembly and Profiling of Candida auris Reveals Novel Insights into Biofilm-Mediated Resistance"

Article Title: Transcriptome Assembly and Profiling of Candida auris Reveals Novel Insights into Biofilm-Mediated Resistance

Journal: mSphere

doi: 10.1128/mSphere.00334-18

Functional annotation of differentially expressed genes reveals upregulation of drug transporters. Gene distribution of significantly upregulated C. auris genes in 24-h biofilms relative to planktonic cells, grouped into biological process (BP), cellular component (CC), and metabolic function (MF) gene ontology categories (A). Log 2 fold change of upregulated ABC and MFS drug transporters within 24-h biofilms (B). All GO terms have a P value of
Figure Legend Snippet: Functional annotation of differentially expressed genes reveals upregulation of drug transporters. Gene distribution of significantly upregulated C. auris genes in 24-h biofilms relative to planktonic cells, grouped into biological process (BP), cellular component (CC), and metabolic function (MF) gene ontology categories (A). Log 2 fold change of upregulated ABC and MFS drug transporters within 24-h biofilms (B). All GO terms have a P value of

Techniques Used: Functional Assay

Formation and development of Candida auris biofilms. Schematic representation of the transcriptional mediators of the three main stages of C. auris biofilm development: adherence of yeast cells to surface (early phase), proliferation (intermediate phase), and maturation into a structured biofilm (mature phase).
Figure Legend Snippet: Formation and development of Candida auris biofilms. Schematic representation of the transcriptional mediators of the three main stages of C. auris biofilm development: adherence of yeast cells to surface (early phase), proliferation (intermediate phase), and maturation into a structured biofilm (mature phase).

Techniques Used:

Candida auris biofilm development correlates with increased antifungal tolerance. Candida auris biofilms were standardized at 1 × 10 6 CFU/ml and grown for 4, 12, and 24 h. Biofilm biomass was then quantified using the crystal violet assay, with the composition of biofilm cells enumerated using qPCR and represented by a box-and-whisker plot as the total biomass of four C. auris isolates (A, left y axis). Planktonic susceptibility testing was performed against serially diluted miconazole, micafungin, and amphotericin B concentrations using the CLSI guidelines, with biofilm susceptibility testing performed using the XTT assay and with median MIC values plotted (A, right y axis). In addition, biofilms were grown, fixed, and processed for SEM before imaging using a JEOL-JSM-6400 scanning electron microscope. Micrographs represent lower magnification (×1,000) and higher magnification (inset, ×5,000) of biofilms grown for 4 h (Bi), 12 h (Bii), and 24 h (Biii).
Figure Legend Snippet: Candida auris biofilm development correlates with increased antifungal tolerance. Candida auris biofilms were standardized at 1 × 10 6 CFU/ml and grown for 4, 12, and 24 h. Biofilm biomass was then quantified using the crystal violet assay, with the composition of biofilm cells enumerated using qPCR and represented by a box-and-whisker plot as the total biomass of four C. auris isolates (A, left y axis). Planktonic susceptibility testing was performed against serially diluted miconazole, micafungin, and amphotericin B concentrations using the CLSI guidelines, with biofilm susceptibility testing performed using the XTT assay and with median MIC values plotted (A, right y axis). In addition, biofilms were grown, fixed, and processed for SEM before imaging using a JEOL-JSM-6400 scanning electron microscope. Micrographs represent lower magnification (×1,000) and higher magnification (inset, ×5,000) of biofilms grown for 4 h (Bi), 12 h (Bii), and 24 h (Biii).

Techniques Used: Crystal Violet Assay, Real-time Polymerase Chain Reaction, Whisker Assay, XTT Assay, Imaging, Microscopy

20) Product Images from "Swarming motility and biofilm formation of Paenibacillus larvae, the etiological agent of American Foulbrood of honey bees (Apis mellifera)"

Article Title: Swarming motility and biofilm formation of Paenibacillus larvae, the etiological agent of American Foulbrood of honey bees (Apis mellifera)

Journal: Scientific Reports

doi: 10.1038/s41598-018-27193-8

Planktonic cells of P. larvae ERIC I (ATCC9545) and P. larvae ERIC II (DSM25430). Bacterial suspensions of P. larvae ERIC I (ATCC9545; A ) and P. larvae ERIC II (DSM25430; B ) in brain heart infusion (BHI) broth were incubated under constant agitation to prevent biofilm formation. Cultures of planktonic cells were stained with Congo red ( A , B ) while BHI medium stained with Congo red ( C ) or without staining ( D ) served as negative controls. Representative pictures are shown.
Figure Legend Snippet: Planktonic cells of P. larvae ERIC I (ATCC9545) and P. larvae ERIC II (DSM25430). Bacterial suspensions of P. larvae ERIC I (ATCC9545; A ) and P. larvae ERIC II (DSM25430; B ) in brain heart infusion (BHI) broth were incubated under constant agitation to prevent biofilm formation. Cultures of planktonic cells were stained with Congo red ( A , B ) while BHI medium stained with Congo red ( C ) or without staining ( D ) served as negative controls. Representative pictures are shown.

Techniques Used: Incubation, Staining

Involvement of paenilarvin in biofilm formation of P. larvae ERIC II. Wild-type P. larvae ERIC II (DSM25430 wt; A , C ) and a corresponding inactivation mutant for the paenilarvin gene cluster (DSM25430 Δ itu ; B , D ) were tested in biofilm assays. Bacterial suspensions in brain heart infusion (BHI) broth were incubated without agitation in six-well-plates at 37 °C for five ( A , B ) and six ( C , D ) days. For both strains, three biological replicates were performed both for obtaining pictures under unstained conditions ( A , B ) and after Congo red staining ( C , D ). Representative pictures are shown. ( E ) Biofilm formation was quantified via determining the amount of Conge red dye retained in the biofilms upon centrifugation. Data represent mean values ± SD of three independent experiments. The difference between the wild-type and corresponding inactivation mutant was not significant (p = 0.9889, Student’s t-test).
Figure Legend Snippet: Involvement of paenilarvin in biofilm formation of P. larvae ERIC II. Wild-type P. larvae ERIC II (DSM25430 wt; A , C ) and a corresponding inactivation mutant for the paenilarvin gene cluster (DSM25430 Δ itu ; B , D ) were tested in biofilm assays. Bacterial suspensions in brain heart infusion (BHI) broth were incubated without agitation in six-well-plates at 37 °C for five ( A , B ) and six ( C , D ) days. For both strains, three biological replicates were performed both for obtaining pictures under unstained conditions ( A , B ) and after Congo red staining ( C , D ). Representative pictures are shown. ( E ) Biofilm formation was quantified via determining the amount of Conge red dye retained in the biofilms upon centrifugation. Data represent mean values ± SD of three independent experiments. The difference between the wild-type and corresponding inactivation mutant was not significant (p = 0.9889, Student’s t-test).

Techniques Used: Mutagenesis, Incubation, Staining, Centrifugation

Biofilm formation of P. larvae ERIC I (ATCC9545) and P. larvae ERIC II (DSM25430) cultivated in static liquid. Bacterial suspensions of P. larvae ERIC I (ATCC9545; A , D , G ) and P. larvae ERIC II (DSM25430; B , E , H ) in brain heart infusion (BHI) broth were incubated without agitation in six-well-plates at 37 °C for five ( A , B ) and six ( D , E ) days, or in 96-well-plates for six days ( G , H ). For each P. larvae genotype, three biological replicates were performed both for obtaining pictures under unstained conditions ( A , B ), after Congo red staining of the extracellular matrix ( D , E ), and Crystal violet staining of the bacterial cells adherent to the well walls ( G , H ). Negative controls for each assay are shown in ( C , F , and I ). Representative pictures are shown.
Figure Legend Snippet: Biofilm formation of P. larvae ERIC I (ATCC9545) and P. larvae ERIC II (DSM25430) cultivated in static liquid. Bacterial suspensions of P. larvae ERIC I (ATCC9545; A , D , G ) and P. larvae ERIC II (DSM25430; B , E , H ) in brain heart infusion (BHI) broth were incubated without agitation in six-well-plates at 37 °C for five ( A , B ) and six ( D , E ) days, or in 96-well-plates for six days ( G , H ). For each P. larvae genotype, three biological replicates were performed both for obtaining pictures under unstained conditions ( A , B ), after Congo red staining of the extracellular matrix ( D , E ), and Crystal violet staining of the bacterial cells adherent to the well walls ( G , H ). Negative controls for each assay are shown in ( C , F , and I ). Representative pictures are shown.

Techniques Used: Incubation, Staining

Fluorescence microscopy of P. larvae floating biofilms. Bacterial suspensions of P. larvae ERIC I (ATCC9545; A , C ) and P. larvae ERIC II (DSM25430; B , D ) in Sf-900 II SFM medium supplemented with 30 µg/ml thioflavin S were incubated without agitation in 96-well-plates at 37 °C for six days. The thioflavin S-stained extracellular matrix in the floating biofilms was visualized using fluorescence microscopy; Z-stack processing was performed to obtain three-dimensional images of the wells containing the biofilms ( A , B ) and the region within the wells where the biofilms were located ( C , D ). Bars represent 20 µm.
Figure Legend Snippet: Fluorescence microscopy of P. larvae floating biofilms. Bacterial suspensions of P. larvae ERIC I (ATCC9545; A , C ) and P. larvae ERIC II (DSM25430; B , D ) in Sf-900 II SFM medium supplemented with 30 µg/ml thioflavin S were incubated without agitation in 96-well-plates at 37 °C for six days. The thioflavin S-stained extracellular matrix in the floating biofilms was visualized using fluorescence microscopy; Z-stack processing was performed to obtain three-dimensional images of the wells containing the biofilms ( A , B ) and the region within the wells where the biofilms were located ( C , D ). Bars represent 20 µm.

Techniques Used: Fluorescence, Microscopy, Incubation, Staining

21) Product Images from "Role of Viscoelasticity in Bacterial Killing by Antimicrobials in Differently Grown Pseudomonas aeruginosa Biofilms"

Article Title: Role of Viscoelasticity in Bacterial Killing by Antimicrobials in Differently Grown Pseudomonas aeruginosa Biofilms

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.01972-18

Microscopic images of P. aeruginosa ATCC 39324 biofilms grown in ASM + , ASM − , and LB medium. (a) Two-dimensional, cross-sectional, OCT images. Scale bars, 200 μm. (b) CLSM two-dimensional overlay (left) and three-dimensional (right) images of SYTO9-stained biofilms, yielding green-fluorescent bacteria. Scale bars, 100 μm.
Figure Legend Snippet: Microscopic images of P. aeruginosa ATCC 39324 biofilms grown in ASM + , ASM − , and LB medium. (a) Two-dimensional, cross-sectional, OCT images. Scale bars, 200 μm. (b) CLSM two-dimensional overlay (left) and three-dimensional (right) images of SYTO9-stained biofilms, yielding green-fluorescent bacteria. Scale bars, 100 μm.

Techniques Used: Confocal Laser Scanning Microscopy, Staining

Numbers of CFU per square centimeter in P. aeruginosa ATCC 39324 biofilms grown in ASM + , ASM − , or LB medium after 24-h exposure to different concentrations of tobramycin, colistin, or the antimicrobial peptide AA-230 in PBS, with PBS as a control. Error bars denote standard deviations for at least nine different biofilms, taken from different pans in three separate CDFF runs. Significant differences ( P
Figure Legend Snippet: Numbers of CFU per square centimeter in P. aeruginosa ATCC 39324 biofilms grown in ASM + , ASM − , or LB medium after 24-h exposure to different concentrations of tobramycin, colistin, or the antimicrobial peptide AA-230 in PBS, with PBS as a control. Error bars denote standard deviations for at least nine different biofilms, taken from different pans in three separate CDFF runs. Significant differences ( P

Techniques Used:

Stress relaxation analysis of P. aeruginosa ATCC 39324 biofilms grown in ASM + , ASM − , or LB medium. (a) Examples of the normalized stress in compressed biofilms (strain of 0.2) as a function of relaxation time. Stress at time zero amounted 2.2 kPa for all biofilms, regardless of the growth medium. (b) Quality of fitting of the stress relaxation data to a generalized Maxwell model as a function of the number of elements included in the model. The quality of the fit is indicated by chi-squared values. (c) Distribution of the relative importance of individual Maxwell elements (three-element model) for differently grown P. aeruginosa biofilms over different relaxation time constant ranges. Each data point represents a single measurement of 30 biofilms, taking 10 biofilms from different pans in three separate CDFF runs. Median values are indicated by horizontal lines. Significant differences ( P
Figure Legend Snippet: Stress relaxation analysis of P. aeruginosa ATCC 39324 biofilms grown in ASM + , ASM − , or LB medium. (a) Examples of the normalized stress in compressed biofilms (strain of 0.2) as a function of relaxation time. Stress at time zero amounted 2.2 kPa for all biofilms, regardless of the growth medium. (b) Quality of fitting of the stress relaxation data to a generalized Maxwell model as a function of the number of elements included in the model. The quality of the fit is indicated by chi-squared values. (c) Distribution of the relative importance of individual Maxwell elements (three-element model) for differently grown P. aeruginosa biofilms over different relaxation time constant ranges. Each data point represents a single measurement of 30 biofilms, taking 10 biofilms from different pans in three separate CDFF runs. Median values are indicated by horizontal lines. Significant differences ( P

Techniques Used:

22) Product Images from "Effect of divalent ions and a polyphosphate on composition, structure, and stiffness of simulated drinking water biofilms"

Article Title: Effect of divalent ions and a polyphosphate on composition, structure, and stiffness of simulated drinking water biofilms

Journal: NPJ Biofilms and Microbiomes

doi: 10.1038/s41522-018-0058-1

a TGA curve showing the percentage of biofilm weight at a certain temperature with respect to the initial biofilm weight as a function of temperature and b derivative thermogravimetric analysis (DTG) curve showing the change of biofilm decomposition rate as a function of temperature. The error bars represent the standard deviation of the measurements for the replicates
Figure Legend Snippet: a TGA curve showing the percentage of biofilm weight at a certain temperature with respect to the initial biofilm weight as a function of temperature and b derivative thermogravimetric analysis (DTG) curve showing the change of biofilm decomposition rate as a function of temperature. The error bars represent the standard deviation of the measurements for the replicates

Techniques Used: Standard Deviation

The frequency distributions of elastic modulus values for a the hard-groundwater biofilms and b soft-groundwater biofilms
Figure Legend Snippet: The frequency distributions of elastic modulus values for a the hard-groundwater biofilms and b soft-groundwater biofilms

Techniques Used:

a Average thickness and b selected OCT images of biofilms developed from hard groundwater, soft groundwater, and groundwater containing SHMP
Figure Legend Snippet: a Average thickness and b selected OCT images of biofilms developed from hard groundwater, soft groundwater, and groundwater containing SHMP

Techniques Used:

ATR-FTIR spectra of hard-groundwater biofilms, soft-groundwater biofilms, and SHMP biofilms. The main absorbance peaks are shown in the figure
Figure Legend Snippet: ATR-FTIR spectra of hard-groundwater biofilms, soft-groundwater biofilms, and SHMP biofilms. The main absorbance peaks are shown in the figure

Techniques Used:

a The selected biofilm oscillation amplitude curves across a frequency ranged from 10 to 500 Hz. The peak in each curve represents the mechanical resonance frequency of each biofilm. b The average resonance frequency and standard deviation for each biofilm sample. The error bars represent the standard deviation of the measurements for the replicates
Figure Legend Snippet: a The selected biofilm oscillation amplitude curves across a frequency ranged from 10 to 500 Hz. The peak in each curve represents the mechanical resonance frequency of each biofilm. b The average resonance frequency and standard deviation for each biofilm sample. The error bars represent the standard deviation of the measurements for the replicates

Techniques Used: Standard Deviation

23) Product Images from "Phenalen-1-one-Mediated Antimicrobial Photodynamic Therapy: Antimicrobial Efficacy in a Periodontal Biofilm Model and Flow Cytometric Evaluation of Cytoplasmic Membrane Damage"

Article Title: Phenalen-1-one-Mediated Antimicrobial Photodynamic Therapy: Antimicrobial Efficacy in a Periodontal Biofilm Model and Flow Cytometric Evaluation of Cytoplasmic Membrane Damage

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2018.00688

Exemplary visualization of polymicrobial biofilms by means of scanning electron microscopy. Exemplary SEM visualization of randomly selected fields of untreated biofilms (PS-L–), biofilms treated with aPDT using SAPYR (SAPYR+L+) or SAGUA (SAGUA+L+) or treated with CHX (CHX 0.2%) in 3,000-fold, 12,000-fold, and 24,000-fold magnification. In the CHX-treated biofilms white arrows show debris most likely originating from killed cells on the top layer of the biofilms.
Figure Legend Snippet: Exemplary visualization of polymicrobial biofilms by means of scanning electron microscopy. Exemplary SEM visualization of randomly selected fields of untreated biofilms (PS-L–), biofilms treated with aPDT using SAPYR (SAPYR+L+) or SAGUA (SAGUA+L+) or treated with CHX (CHX 0.2%) in 3,000-fold, 12,000-fold, and 24,000-fold magnification. In the CHX-treated biofilms white arrows show debris most likely originating from killed cells on the top layer of the biofilms.

Techniques Used: Electron Microscopy

Spectroscopic measurements for release of nucleic acids. OD medians, 1st and 3rd quartiles of the supernatants of biofilms treated with phenalen-1-one mediated aPDT (groups: PS-L–, PS-L+, SAPYR+L-, SAPYR+L+, SAGUA+L-, and SAGUA+L+) or positive control (lysozyme treatment followed by Proteinase K digestion), as measured at 260 nm for release of nucleic acids.
Figure Legend Snippet: Spectroscopic measurements for release of nucleic acids. OD medians, 1st and 3rd quartiles of the supernatants of biofilms treated with phenalen-1-one mediated aPDT (groups: PS-L–, PS-L+, SAPYR+L-, SAPYR+L+, SAGUA+L-, and SAGUA+L+) or positive control (lysozyme treatment followed by Proteinase K digestion), as measured at 260 nm for release of nucleic acids.

Techniques Used: Positive Control

24) Product Images from "Regulation of Monospecies and Mixed Biofilms Formation of Skin Staphylococcus aureus and Cutibacterium acnes by Human Natriuretic Peptides"

Article Title: Regulation of Monospecies and Mixed Biofilms Formation of Skin Staphylococcus aureus and Cutibacterium acnes by Human Natriuretic Peptides

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2018.02912

Parameters of monospecies biofilms of S. aureus MFP03 measured by CLSM in different cultivation conditions. (A) Average biofilm biomass density and (B) Average biofilm thickness. ∗ P
Figure Legend Snippet: Parameters of monospecies biofilms of S. aureus MFP03 measured by CLSM in different cultivation conditions. (A) Average biofilm biomass density and (B) Average biofilm thickness. ∗ P

Techniques Used: Confocal Laser Scanning Microscopy

Contribution of C. acnes RT5 and S. aureus MFP03 to binary biofilms formed in the absence or presence of ANP or CNP. (A) Simultaneously grown biofilms; (B) Biofilms based on pre-formed S. aureus MFP03 biofilms; (C) Biofilms, based on pre-formed C. acnes RT5. ∗ P
Figure Legend Snippet: Contribution of C. acnes RT5 and S. aureus MFP03 to binary biofilms formed in the absence or presence of ANP or CNP. (A) Simultaneously grown biofilms; (B) Biofilms based on pre-formed S. aureus MFP03 biofilms; (C) Biofilms, based on pre-formed C. acnes RT5. ∗ P

Techniques Used: Aqueous Normal-phase Chromatography

Parameters of mixed biofilms of S. aureus MFP03 and C. acnes strains at 37°C. (A) S. aureus MFP03 CFU and (B) MTT biofilm values. ∗ P
Figure Legend Snippet: Parameters of mixed biofilms of S. aureus MFP03 and C. acnes strains at 37°C. (A) S. aureus MFP03 CFU and (B) MTT biofilm values. ∗ P

Techniques Used: MTT Assay

Parameters of monospecies biofilms of C. acnes measured by CLSM in different cultivation conditions. (A) Average C. acnes RT4 biofilm biomass density; (B) Average C. acnes RT5 biofilm biomass density; (C) Average C. acnes RT4 biofilm thickness; (D) Average C. acnes RT5 biofilm thickness. ∗ P
Figure Legend Snippet: Parameters of monospecies biofilms of C. acnes measured by CLSM in different cultivation conditions. (A) Average C. acnes RT4 biofilm biomass density; (B) Average C. acnes RT5 biofilm biomass density; (C) Average C. acnes RT4 biofilm thickness; (D) Average C. acnes RT5 biofilm thickness. ∗ P

Techniques Used: Confocal Laser Scanning Microscopy

Effects of ANP and CNP on mixed biofilms of S. aureus MFP03 and C. acnes RT5 at 33°C. (A) S. aureus MFP03 CFU and (B) MTT biofilm values. ∗ P
Figure Legend Snippet: Effects of ANP and CNP on mixed biofilms of S. aureus MFP03 and C. acnes RT5 at 33°C. (A) S. aureus MFP03 CFU and (B) MTT biofilm values. ∗ P

Techniques Used: Aqueous Normal-phase Chromatography, MTT Assay

CLSM images of FISH-labeled mixed biofilms of S. aureus MFP03 and C. acnes RT5. (A) Simultaneously grown biofilm, Control; (B) Simultaneously grown biofilm, ANP 10 -8 M; (C) Simultaneously grown biofilm, CNP 10 -7 M; (D) Biofilm based on pre-formed S. aureus MFP03 biofilm, Control; (E) Biofilm based on pre-formed S. aureus MFP03 biofilm, ANP 10 -8 M; (F) Biofilm based on pre-formed S. aureus MFP03 biofilm, CNP 10 -7 M; (G) Biofilm based on pre-formed C. acnes RT5 biofilm, Control; (H) Biofilm based on pre-formed C. acnes RT5 biofilm, ANP 10 -8 M; (I) Biofilm based on pre-formed C. acnes RT5 biofilm, CNP 10 -7 M. Pink, C. acnes RT5 FISH labeling; Blue, DAPI staining (total biomass).
Figure Legend Snippet: CLSM images of FISH-labeled mixed biofilms of S. aureus MFP03 and C. acnes RT5. (A) Simultaneously grown biofilm, Control; (B) Simultaneously grown biofilm, ANP 10 -8 M; (C) Simultaneously grown biofilm, CNP 10 -7 M; (D) Biofilm based on pre-formed S. aureus MFP03 biofilm, Control; (E) Biofilm based on pre-formed S. aureus MFP03 biofilm, ANP 10 -8 M; (F) Biofilm based on pre-formed S. aureus MFP03 biofilm, CNP 10 -7 M; (G) Biofilm based on pre-formed C. acnes RT5 biofilm, Control; (H) Biofilm based on pre-formed C. acnes RT5 biofilm, ANP 10 -8 M; (I) Biofilm based on pre-formed C. acnes RT5 biofilm, CNP 10 -7 M. Pink, C. acnes RT5 FISH labeling; Blue, DAPI staining (total biomass).

Techniques Used: Confocal Laser Scanning Microscopy, Fluorescence In Situ Hybridization, Labeling, Aqueous Normal-phase Chromatography, Staining

25) Product Images from "Inhibitory effects of 7-epiclusianone on glucan synthesis, acidogenicity and biofilm formation by Streptococcus mutans"

Article Title: Inhibitory effects of 7-epiclusianone on glucan synthesis, acidogenicity and biofilm formation by Streptococcus mutans

Journal:

doi: 10.1111/j.1574-6968.2008.01117.x

Influence of 7-epiclusianone on the pH values in the culture medium during Streptococcus mutans biofilm formation. The medium was replaced daily with fresh medium. The pH values ( n = 9) were determined after 4, 8, 12 and 24 h of incubation.
Figure Legend Snippet: Influence of 7-epiclusianone on the pH values in the culture medium during Streptococcus mutans biofilm formation. The medium was replaced daily with fresh medium. The pH values ( n = 9) were determined after 4, 8, 12 and 24 h of incubation.

Techniques Used: Incubation

26) Product Images from "Proteomic Analysis of Campylobacter jejuni 11168 Biofilms Reveals a Role for the Motility Complex in Biofilm Formation"

Article Title: Proteomic Analysis of Campylobacter jejuni 11168 Biofilms Reveals a Role for the Motility Complex in Biofilm Formation

Journal:

doi: 10.1128/JB.01975-05

SEM of C. jejuni 11168 attached to various surfaces. (A) Biofilm formed on stainless steel, with the inset demonstrating extracellular fiber-like material. (B) Biofilm formed on nitrocellulose. (C) Larger image of biofilm formed on nitrocellulose, demonstrating
Figure Legend Snippet: SEM of C. jejuni 11168 attached to various surfaces. (A) Biofilm formed on stainless steel, with the inset demonstrating extracellular fiber-like material. (B) Biofilm formed on nitrocellulose. (C) Larger image of biofilm formed on nitrocellulose, demonstrating

Techniques Used:

Expansion of a second set of two-dimensional gel images of C. jejuni planktonic (A) and biofilm (B) cell lysates showing additional flagellar proteins in the enlarged pH 4 to 6 range. Proteins overexpressed in biofilm cells, indicated by arrows, were
Figure Legend Snippet: Expansion of a second set of two-dimensional gel images of C. jejuni planktonic (A) and biofilm (B) cell lysates showing additional flagellar proteins in the enlarged pH 4 to 6 range. Proteins overexpressed in biofilm cells, indicated by arrows, were

Techniques Used: Two-Dimensional Gel Electrophoresis

27) Product Images from "Eradication of Pseudomonas aeruginosa biofilms and persister cells using an electrochemical scaffold and enhanced antibiotic susceptibility"

Article Title: Eradication of Pseudomonas aeruginosa biofilms and persister cells using an electrochemical scaffold and enhanced antibiotic susceptibility

Journal: NPJ Biofilms and Microbiomes

doi: 10.1038/s41522-016-0003-0

a Schematic of experimental setup for the treatment of biofilm exposed to an e-scaffold with an illustration of electrochemical H 2 O 2 production. The electrodes are connected to a potentiostat (not shown in figure). Scientific Reports, 2015. 5, 14908. DOI: 10.1038/srep14908. © Creative Commons Attribution 4.0 International License. b Recovered biofilm cells treated with e-scaffold and tobramycin. c Treatment of biofilms and persister cells with e-scaffold
Figure Legend Snippet: a Schematic of experimental setup for the treatment of biofilm exposed to an e-scaffold with an illustration of electrochemical H 2 O 2 production. The electrodes are connected to a potentiostat (not shown in figure). Scientific Reports, 2015. 5, 14908. DOI: 10.1038/srep14908. © Creative Commons Attribution 4.0 International License. b Recovered biofilm cells treated with e-scaffold and tobramycin. c Treatment of biofilms and persister cells with e-scaffold

Techniques Used:

E-scaffold enhances tobramycin susceptibility in P. aeruginosa PAO1 biofilms. Bars represent means of logarithms of colony-forming units of viable biofilm cells. Error bars represent the standard error of the means from three biological replicates. The symbols *, **, ***, and **** represent significant differences in tobramycin susceptibility between e-scaffold treated biofilms + tobramycin and untreated biofilms + tobramycin ( n = 3, *, P = 0.002 ; **, ***, **** P ≤ 0.001; paired t -test)
Figure Legend Snippet: E-scaffold enhances tobramycin susceptibility in P. aeruginosa PAO1 biofilms. Bars represent means of logarithms of colony-forming units of viable biofilm cells. Error bars represent the standard error of the means from three biological replicates. The symbols *, **, ***, and **** represent significant differences in tobramycin susceptibility between e-scaffold treated biofilms + tobramycin and untreated biofilms + tobramycin ( n = 3, *, P = 0.002 ; **, ***, **** P ≤ 0.001; paired t -test)

Techniques Used:

Exposure to e-scaffold eradicates the persister cells in P. aeruginosa PAO1 biofilms. Bars represent means of logarithms of colony-forming units of viable biofilm cells. Error bars represent the standard errors of the means from three biological replicates. The symbol * denotes a significant difference compared to total biofilm cells ( n = 3, P
Figure Legend Snippet: Exposure to e-scaffold eradicates the persister cells in P. aeruginosa PAO1 biofilms. Bars represent means of logarithms of colony-forming units of viable biofilm cells. Error bars represent the standard errors of the means from three biological replicates. The symbol * denotes a significant difference compared to total biofilm cells ( n = 3, P

Techniques Used:

Scanning electron microscopy (SEM) images showing untreated, e-scaffold treated and exogenous H 2 O 2 treated P. aeruginosa PAO1 biofilm cells. Three representative images are shown for each treatment. Red arrows indicate cells showing a stressed membrane
Figure Legend Snippet: Scanning electron microscopy (SEM) images showing untreated, e-scaffold treated and exogenous H 2 O 2 treated P. aeruginosa PAO1 biofilm cells. Three representative images are shown for each treatment. Red arrows indicate cells showing a stressed membrane

Techniques Used: Electron Microscopy

E-scaffold increases OH• formation and membrane permeability. a Increase in fluorescence of HPF-stained e-scaffold treated and exogenous H 2 O 2 treated P. aeruginosa PAO1 biofilm cells indicates increased OH• formation compared to untreated biofilms. b Increase in fluorescence of propidium iodide (PI) indicates increased membrane permeability of P. aeruginosa PAO1 cells after exposure to e-scaffold. c Increase in fluorescence of 3, 3′-dipropylthiacarbocyanine iodide, a membrane potential sensitive dye, in e-scaffold treated P. aeruginosa PAO1 cells verifies bacterial membrane depolarization by e-scaffold resulting in increased membrane permeability. Error bars represent standard errors of means for at least three biological replicates. The symbol * indicates a significant difference from the untreated biofilm cells ( n = 3, P
Figure Legend Snippet: E-scaffold increases OH• formation and membrane permeability. a Increase in fluorescence of HPF-stained e-scaffold treated and exogenous H 2 O 2 treated P. aeruginosa PAO1 biofilm cells indicates increased OH• formation compared to untreated biofilms. b Increase in fluorescence of propidium iodide (PI) indicates increased membrane permeability of P. aeruginosa PAO1 cells after exposure to e-scaffold. c Increase in fluorescence of 3, 3′-dipropylthiacarbocyanine iodide, a membrane potential sensitive dye, in e-scaffold treated P. aeruginosa PAO1 cells verifies bacterial membrane depolarization by e-scaffold resulting in increased membrane permeability. Error bars represent standard errors of means for at least three biological replicates. The symbol * indicates a significant difference from the untreated biofilm cells ( n = 3, P

Techniques Used: Permeability, Fluorescence, Staining

28) Product Images from "Activity of Antibiotics against Staphylococcus aureus in an In Vitro Model of Biofilms in the Context of Cystic Fibrosis: Influence of the Culture Medium"

Article Title: Activity of Antibiotics against Staphylococcus aureus in an In Vitro Model of Biofilms in the Context of Cystic Fibrosis: Influence of the Culture Medium

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.00602-19

Comparison of antibiotic maximal efficacies ( E max ) expressed as the reduction in the number of CFU from that for the control (left) or as a percentage of the reduction in metabolic activity (resorufin fluorescence; middle) or biofilm mass (crystal violet [CV] absorbance; right) compared to that for an untreated biofilm for strain ATCC 25923 (open bar) or ATCC 33591 (closed bars) grown in Trypticase soy broth supplemented with 1% glucose and 2% NaCl (TGN) or in artificial sputum medium (ASM). MEM, meropenem; VAN, vancomycin; LZD, linezolid; AZM, azithromycin; RIF, rifampin; CIP, ciprofloxacin; TOB, tobramycin. Values are means ± SEM. Statistical analyses were by one-way analysis of variance (ANOVA) with Tukey’s posttest for multiple comparisons; values with different letters are significantly different from each other ( P  
Figure Legend Snippet: Comparison of antibiotic maximal efficacies ( E max ) expressed as the reduction in the number of CFU from that for the control (left) or as a percentage of the reduction in metabolic activity (resorufin fluorescence; middle) or biofilm mass (crystal violet [CV] absorbance; right) compared to that for an untreated biofilm for strain ATCC 25923 (open bar) or ATCC 33591 (closed bars) grown in Trypticase soy broth supplemented with 1% glucose and 2% NaCl (TGN) or in artificial sputum medium (ASM). MEM, meropenem; VAN, vancomycin; LZD, linezolid; AZM, azithromycin; RIF, rifampin; CIP, ciprofloxacin; TOB, tobramycin. Values are means ± SEM. Statistical analyses were by one-way analysis of variance (ANOVA) with Tukey’s posttest for multiple comparisons; values with different letters are significantly different from each other ( P  

Techniques Used: Activity Assay, Fluorescence

Metabolic activity in planktonic cultures or biofilm cultures grown in Trypticase soy broth supplemented with 1% glucose and 2% NaCl (TGN) or in artificial sputum medium (ASM). (Left) Resorufin fluorescence signal recorded after 30 min of incubation of planktonic bacteria at increasing inocula with 10-mg/liter resazurin. (Right) Resorufin fluorescence signal recorded after 30 min of incubation of a 24-h-old biofilm with 10-mg/liter resazurin. Data are means ± SD for triplicates in a single experiment or means ± SEM from at least 3 independent experiments performed in triplicate. Statistical analyses comparing strains in each individual medium were performed by one-way analysis of variance (ANOVA) with Tukey’s posttest for multiple comparisons; values with different letters are significantly different from each other ( P  
Figure Legend Snippet: Metabolic activity in planktonic cultures or biofilm cultures grown in Trypticase soy broth supplemented with 1% glucose and 2% NaCl (TGN) or in artificial sputum medium (ASM). (Left) Resorufin fluorescence signal recorded after 30 min of incubation of planktonic bacteria at increasing inocula with 10-mg/liter resazurin. (Right) Resorufin fluorescence signal recorded after 30 min of incubation of a 24-h-old biofilm with 10-mg/liter resazurin. Data are means ± SD for triplicates in a single experiment or means ± SEM from at least 3 independent experiments performed in triplicate. Statistical analyses comparing strains in each individual medium were performed by one-way analysis of variance (ANOVA) with Tukey’s posttest for multiple comparisons; values with different letters are significantly different from each other ( P  

Techniques Used: Activity Assay, Fluorescence, Incubation

Morphology and counts of colonies from biofilms of ATCC 25923 cultivated in artificial sputum medium (ASM) and exposed for 24 h to tobramycin (TOB) at 5 (TOB 5), 10 (TOB 10), or 100 (TOB 100) mg/liter or under control conditions (no antibiotic added as a control [CT]). Samples were plated on Columbia blood agar, TSA, or TSA supplemented with either 1 mg/liter hemin (TSAH), menadione (TSAM). or both hemin and menadione (TSAMH). Yellow arrows indicate a typical small colony.
Figure Legend Snippet: Morphology and counts of colonies from biofilms of ATCC 25923 cultivated in artificial sputum medium (ASM) and exposed for 24 h to tobramycin (TOB) at 5 (TOB 5), 10 (TOB 10), or 100 (TOB 100) mg/liter or under control conditions (no antibiotic added as a control [CT]). Samples were plated on Columbia blood agar, TSA, or TSA supplemented with either 1 mg/liter hemin (TSAH), menadione (TSAM). or both hemin and menadione (TSAMH). Yellow arrows indicate a typical small colony.

Techniques Used:

29) Product Images from "Intercellular adhesion and biocide resistance in nontypeable Haemophilus influenzae biofilms"

Article Title: Intercellular adhesion and biocide resistance in nontypeable Haemophilus influenzae biofilms

Journal: Microbial pathogenesis

doi: 10.1016/j.micpath.2009.01.004

DNase I and proteinase K sensitize NTHi NJ9725 biofilms to killing by CPC. (A) Biofilms cultured for 24 in microtiter plates were rinsed with water and treated for 10 min with DNase I buffer (mock pre-treatment) or DNase I buffer containing 1 mg mL -1
Figure Legend Snippet: DNase I and proteinase K sensitize NTHi NJ9725 biofilms to killing by CPC. (A) Biofilms cultured for 24 in microtiter plates were rinsed with water and treated for 10 min with DNase I buffer (mock pre-treatment) or DNase I buffer containing 1 mg mL -1

Techniques Used: Cell Culture

DNase I and proteinase K inhibit NTHi biofilm formation. (A) NTHi strain NJ9725 was cultured in polystyrene tubes in BHI broth or BHI broth supplemented with 1 mg mL -1 of DNase I or proteinase K. After increasing amounts of time, tubes were washed with
Figure Legend Snippet: DNase I and proteinase K inhibit NTHi biofilm formation. (A) NTHi strain NJ9725 was cultured in polystyrene tubes in BHI broth or BHI broth supplemented with 1 mg mL -1 of DNase I or proteinase K. After increasing amounts of time, tubes were washed with

Techniques Used: Cell Culture

Detachment of 24-h-old NTHi biofilms by DNase I and proteinase K. All enzymes were tested in enzyme buffer at a concentration of 1 mg mL -1 . (A) Detachment of biofilms in 1 h by proteinase K. (B) Time course for proteinase K-induced detachment of NJ9725
Figure Legend Snippet: Detachment of 24-h-old NTHi biofilms by DNase I and proteinase K. All enzymes were tested in enzyme buffer at a concentration of 1 mg mL -1 . (A) Detachment of biofilms in 1 h by proteinase K. (B) Time course for proteinase K-induced detachment of NJ9725

Techniques Used: Concentration Assay

Fluid convection through 24-h-old NTHi NJ9725 biofilms cultured in centrifugal filter devices. Biofilms were cultured in BHI broth or BHI broth supplemented with 1 mg mL -1 DNase I or proteinase K. The devices were subjected to low-speed centrifugation
Figure Legend Snippet: Fluid convection through 24-h-old NTHi NJ9725 biofilms cultured in centrifugal filter devices. Biofilms were cultured in BHI broth or BHI broth supplemented with 1 mg mL -1 DNase I or proteinase K. The devices were subjected to low-speed centrifugation

Techniques Used: Convection, Cell Culture, Centrifugation

NTHi biofilms exhibit increased resistance to killing by detergents, antiseptics and disinfectants. NTHI NJ9725 biofilms cultured for 24 h in 12-well microtiter plates (left panel), or biofilm cells detached from the surface and dispersed by vortex agitation
Figure Legend Snippet: NTHi biofilms exhibit increased resistance to killing by detergents, antiseptics and disinfectants. NTHI NJ9725 biofilms cultured for 24 h in 12-well microtiter plates (left panel), or biofilm cells detached from the surface and dispersed by vortex agitation

Techniques Used: Cell Culture

30) Product Images from "Bactericidal Effect of a Photoresponsive Carbon Monoxide-Releasing Nonwoven against Staphylococcus aureus Biofilms"

Article Title: Bactericidal Effect of a Photoresponsive Carbon Monoxide-Releasing Nonwoven against Staphylococcus aureus Biofilms

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.00703-16

Live/dead-stained MRSA biofilms 60 min and 135 min after irradiation at 405 nm. Biofilms were grown on glass (control), PLA, or CORM-1–PLA20. Viable bacteria are shown in green and dead bacteria in red. The CLSM images were taken close (2-μm
Figure Legend Snippet: Live/dead-stained MRSA biofilms 60 min and 135 min after irradiation at 405 nm. Biofilms were grown on glass (control), PLA, or CORM-1–PLA20. Viable bacteria are shown in green and dead bacteria in red. The CLSM images were taken close (2-μm

Techniques Used: Staining, Irradiation, Proximity Ligation Assay, Confocal Laser Scanning Microscopy

Changes in the biofilms in response to irradiation at 405 nm and CO release. Biofilms were grown in the presence (CORM-1–PLA20) or absence (PLA) of CORM-1 or without the nonwoven (glass). (A and B) Quantification of viable cells after irradiation
Figure Legend Snippet: Changes in the biofilms in response to irradiation at 405 nm and CO release. Biofilms were grown in the presence (CORM-1–PLA20) or absence (PLA) of CORM-1 or without the nonwoven (glass). (A and B) Quantification of viable cells after irradiation

Techniques Used: Irradiation, Proximity Ligation Assay

31) Product Images from "Identification and Characterization of a Vibrio cholerae Gene, mbaA, Involved in Maintenance of Biofilm Architecture"

Article Title: Identification and Characterization of a Vibrio cholerae Gene, mbaA, Involved in Maintenance of Biofilm Architecture

Journal: Journal of Bacteriology

doi: 10.1128/JB.185.4.1384-1390.2003

The absence of mbaA leads to accumulation of a larger amount of biofilm matrix material. WT and Δ mbaA mutant strains of V . cholerae N16961 were incubated with borosilicate coverslips during 65 h in LB medium at room temperature. Biofilms were observed after fixation by SEM. The extracellular matrix of the biofilms appears on the micrographs as a white fibrous network among cells. Bar, 1 μm.
Figure Legend Snippet: The absence of mbaA leads to accumulation of a larger amount of biofilm matrix material. WT and Δ mbaA mutant strains of V . cholerae N16961 were incubated with borosilicate coverslips during 65 h in LB medium at room temperature. Biofilms were observed after fixation by SEM. The extracellular matrix of the biofilms appears on the micrographs as a white fibrous network among cells. Bar, 1 μm.

Techniques Used: Mutagenesis, Incubation

Flagella and MSHA pili are required for biofilm formation in the absence of mbaA . Biofilm development of the indicated V . cholerae strains was followed in LB medium at room temperature over the course of 65 h. Cells that attached to borosilicate tubes after various periods of incubation were stained with crystal violet. The OD 570 was measured to quantify the amount of DMSO-solubilized dye. All assays were performed in triplicates. Shown are the biofilm developments of the WT (closed circles) and Δ mbaA (open circles) strains (A) and those of the Δ mshA (closed triangles) and Δ mbaA- Δ mshA (open triangles) mutant strains (B).
Figure Legend Snippet: Flagella and MSHA pili are required for biofilm formation in the absence of mbaA . Biofilm development of the indicated V . cholerae strains was followed in LB medium at room temperature over the course of 65 h. Cells that attached to borosilicate tubes after various periods of incubation were stained with crystal violet. The OD 570 was measured to quantify the amount of DMSO-solubilized dye. All assays were performed in triplicates. Shown are the biofilm developments of the WT (closed circles) and Δ mbaA (open circles) strains (A) and those of the Δ mshA (closed triangles) and Δ mbaA- Δ mshA (open triangles) mutant strains (B).

Techniques Used: Incubation, Staining, Mutagenesis

Disruption of a novel gene named mbaA in V . cholerae N16961 leads to increased biofilm formation. (A) Crystal violet-stained borosilicate tubes and quantification of the DMSO-solubilized dye are shown for the indicated strains after 30 h of growth in LB broth at room temperature. All assays were performed in triplicates. (B) Genetic organization of the mbaA region on the chromosome of V . cholerae N16961. The a , b , mbaA , c , and d open reading frames correspond to loci VC0705, VC0704, VC0703, VC0702, and VC0701, respectively, based on the DNA sequences obtained from The Institute for Genomic Research microbial genome database. Open reading frames a , mbaA , and c might constitute an operon. The Tn 10 insertion in mbaA and the extent of the in-frame deletion created in mbaA are indicated.
Figure Legend Snippet: Disruption of a novel gene named mbaA in V . cholerae N16961 leads to increased biofilm formation. (A) Crystal violet-stained borosilicate tubes and quantification of the DMSO-solubilized dye are shown for the indicated strains after 30 h of growth in LB broth at room temperature. All assays were performed in triplicates. (B) Genetic organization of the mbaA region on the chromosome of V . cholerae N16961. The a , b , mbaA , c , and d open reading frames correspond to loci VC0705, VC0704, VC0703, VC0702, and VC0701, respectively, based on the DNA sequences obtained from The Institute for Genomic Research microbial genome database. Open reading frames a , mbaA , and c might constitute an operon. The Tn 10 insertion in mbaA and the extent of the in-frame deletion created in mbaA are indicated.

Techniques Used: Staining

The production of EPS is required for biofilm formation in the absence of mbaA . WT, vsp59 :: pGp704 , and Δ mbaA-vps59 :: pGp704 mutant cells of V . cholerae N16961 were incubated in LB broth at room temperature for 40 h. Phase contrast micrographs of biofilms formed on borosilicate coverslips are shown in the left panels. Crystal violet-stained borosilicate tubes are shown in the right panels.
Figure Legend Snippet: The production of EPS is required for biofilm formation in the absence of mbaA . WT, vsp59 :: pGp704 , and Δ mbaA-vps59 :: pGp704 mutant cells of V . cholerae N16961 were incubated in LB broth at room temperature for 40 h. Phase contrast micrographs of biofilms formed on borosilicate coverslips are shown in the left panels. Crystal violet-stained borosilicate tubes are shown in the right panels.

Techniques Used: Mutagenesis, Incubation, Staining

The absence of mbaA leads to alterations in the biofilm architecture. WT and gfp -tagged Δ mbaA mutant cells of V . cholerae N16961 were incubated with borosilicate coverslips in LB broth at room temperature. Biofilm formation on the coverslips was observed at the indicated times. Shown are phase contrast micrographs of biofilms formed after 8 (A) and 40 h (B) and confocal scanning laser micrographs of biofilms formed after 65 h of incubation (C). Micrographs represent optical sections in the x-z plane, with the substratum located at the bottom. Bar, 5 μm.
Figure Legend Snippet: The absence of mbaA leads to alterations in the biofilm architecture. WT and gfp -tagged Δ mbaA mutant cells of V . cholerae N16961 were incubated with borosilicate coverslips in LB broth at room temperature. Biofilm formation on the coverslips was observed at the indicated times. Shown are phase contrast micrographs of biofilms formed after 8 (A) and 40 h (B) and confocal scanning laser micrographs of biofilms formed after 65 h of incubation (C). Micrographs represent optical sections in the x-z plane, with the substratum located at the bottom. Bar, 5 μm.

Techniques Used: Mutagenesis, Incubation

32) Product Images from "Opaque cells signal white cells to form biofilms in Candida albicans"

Article Title: Opaque cells signal white cells to form biofilms in Candida albicans

Journal: The EMBO Journal

doi: 10.1038/sj.emboj.7601099

Developing the hypothesis that opaque cells signal white cells of opposite mating type, through the release of pheromone, to form a biofilm that protects the opposing spatial gradients of pheromone that must develop for chemotropism to proceed. ( A ) In overlapping populations of natural white a / a and white α/α cells in the host, the predominant phenotype is white, but there are rare and transient switches from white to opaque. ( B ) An opaque a / a cell releases a -pheromone, which forms a spatial gradient that induces shmooing and promotes chemotropism of an opaque α/α cell, and vice versa . The spatial gradients of opposing pheromones are diagrammed between an opaque a / a and opaque α/α cell both as a graph of concentration as a function of distance (green and red, respectively) and as a sequence of parallel lines decreasing in length as concentration decreases. ( C ) The a -pheromone released by the rare opaque a / a cell and the α-pheromone released by the rare opaque α/α cell promote biofilm formation in white cells of opposite mating type. The spatial gradients of opposing pheromones are diagrammed as rings separated by greater distances denoting concentration decreases. Small arrows represent pheromone induction of white cells of opposite mating type to join in biofilm formation.wh, white; op, opaque.
Figure Legend Snippet: Developing the hypothesis that opaque cells signal white cells of opposite mating type, through the release of pheromone, to form a biofilm that protects the opposing spatial gradients of pheromone that must develop for chemotropism to proceed. ( A ) In overlapping populations of natural white a / a and white α/α cells in the host, the predominant phenotype is white, but there are rare and transient switches from white to opaque. ( B ) An opaque a / a cell releases a -pheromone, which forms a spatial gradient that induces shmooing and promotes chemotropism of an opaque α/α cell, and vice versa . The spatial gradients of opposing pheromones are diagrammed between an opaque a / a and opaque α/α cell both as a graph of concentration as a function of distance (green and red, respectively) and as a sequence of parallel lines decreasing in length as concentration decreases. ( C ) The a -pheromone released by the rare opaque a / a cell and the α-pheromone released by the rare opaque α/α cell promote biofilm formation in white cells of opposite mating type. The spatial gradients of opposing pheromones are diagrammed as rings separated by greater distances denoting concentration decreases. Small arrows represent pheromone induction of white cells of opposite mating type to join in biofilm formation.wh, white; op, opaque.

Techniques Used: Concentration Assay, Sequencing

A minority of opaque cells enhance white cell biofilm development. Different proportions of white and opaque cells were mixed and the same total number of cells spread on a silicon elastomer. After 90 min of undisturbed incubation, the elastomers were gently rinsed, then flooded with fresh growth medium and gently rocked for 48 h. In all combinations, white cells were 50% a / a and 50% α/α, and opaque cells 50% a / a and 50% α/α. In all combinations, a monolayer of cells covered the elastomer at the initiation of rocking which reflected the proportions of white and opaque cells initially inoculated. ( A, D, G, J ) Z-series projection of multiphoton LSCM scans through the biofilm. ( B, E, H, K ) Single optical section in the middle of the biofilm. ( C, F, I, L ) Z-series projections viewed from the side (90° tilt). At the end of the incubation period, the top of each gel was identified by a precipitous decrease in pixel intensity. The small solid arrows in (B), (H) and (K) point to septae in hyphae. The unfilled arrowhead in (H) points to a conjugation tube. ‘R's in (E) refer to apical reversion to the budding growth form at the ends of conjugation tubes that have failed to fuse. Scale bar in the first image for each horizontal row represents 10 μm.
Figure Legend Snippet: A minority of opaque cells enhance white cell biofilm development. Different proportions of white and opaque cells were mixed and the same total number of cells spread on a silicon elastomer. After 90 min of undisturbed incubation, the elastomers were gently rinsed, then flooded with fresh growth medium and gently rocked for 48 h. In all combinations, white cells were 50% a / a and 50% α/α, and opaque cells 50% a / a and 50% α/α. In all combinations, a monolayer of cells covered the elastomer at the initiation of rocking which reflected the proportions of white and opaque cells initially inoculated. ( A, D, G, J ) Z-series projection of multiphoton LSCM scans through the biofilm. ( B, E, H, K ) Single optical section in the middle of the biofilm. ( C, F, I, L ) Z-series projections viewed from the side (90° tilt). At the end of the incubation period, the top of each gel was identified by a precipitous decrease in pixel intensity. The small solid arrows in (B), (H) and (K) point to septae in hyphae. The unfilled arrowhead in (H) points to a conjugation tube. ‘R's in (E) refer to apical reversion to the budding growth form at the ends of conjugation tubes that have failed to fuse. Scale bar in the first image for each horizontal row represents 10 μm.

Techniques Used: Incubation, Conjugation Assay

33) Product Images from "Fibrinogen-Induced Streptococcus mutans Biofilm Formation and Adherence to Endothelial Cells"

Article Title: Fibrinogen-Induced Streptococcus mutans Biofilm Formation and Adherence to Endothelial Cells

Journal: BioMed Research International

doi: 10.1155/2013/431465

Scanning electron micrographs of S. mutans biofilms formed on plastic tissue culture coverslips following growth in THB-HK medium (a), 50% bovine plasma (b), THB-HK medium + fibrinogen (0.156 mg mL −1 ) (c), and THB-HK medium + sucrose (0.25%) (d). Magnification 1000x.
Figure Legend Snippet: Scanning electron micrographs of S. mutans biofilms formed on plastic tissue culture coverslips following growth in THB-HK medium (a), 50% bovine plasma (b), THB-HK medium + fibrinogen (0.156 mg mL −1 ) (c), and THB-HK medium + sucrose (0.25%) (d). Magnification 1000x.

Techniques Used:

Effect of various concentrations of sucrose added to THB-HK medium in the presence and absence of fibrinogen on biofilm formation by S. mutans . Results are expressed as means ± standard deviations of triplicate assays from two independent experiments. * P
Figure Legend Snippet: Effect of various concentrations of sucrose added to THB-HK medium in the presence and absence of fibrinogen on biofilm formation by S. mutans . Results are expressed as means ± standard deviations of triplicate assays from two independent experiments. * P

Techniques Used:

Effect of various plasma proteins added to THB-HK medium on biofilm formation by S. mutans . Results are expressed as means ± standard deviations of triplicate assays from two independent experiments. * P
Figure Legend Snippet: Effect of various plasma proteins added to THB-HK medium on biofilm formation by S. mutans . Results are expressed as means ± standard deviations of triplicate assays from two independent experiments. * P

Techniques Used:

Effect of various concentrations of human fibrinogen added to THB-HK medium on biofilm formation by S. mutans . Results are expressed as means ± standard deviations of triplicate assays from two independent experiments. * P
Figure Legend Snippet: Effect of various concentrations of human fibrinogen added to THB-HK medium on biofilm formation by S. mutans . Results are expressed as means ± standard deviations of triplicate assays from two independent experiments. * P

Techniques Used:

Biofilm formation by S. mutans following growth in bovine plasma as quantified by crystal violet staining. Results are expressed as means ± standard deviations of triplicate assays from two independent experiments. * P
Figure Legend Snippet: Biofilm formation by S. mutans following growth in bovine plasma as quantified by crystal violet staining. Results are expressed as means ± standard deviations of triplicate assays from two independent experiments. * P

Techniques Used: Staining

34) Product Images from "Terrein is an inhibitor of quorum sensing and c-di-GMP in Pseudomonas aeruginosa: a connection between quorum sensing and c-di-GMP"

Article Title: Terrein is an inhibitor of quorum sensing and c-di-GMP in Pseudomonas aeruginosa: a connection between quorum sensing and c-di-GMP

Journal: Scientific Reports

doi: 10.1038/s41598-018-26974-5

Effects of exogenous QS ligands on the inhibition of biofilm and cellular c-di-GMP by terrein. ( a and b ) Biofilm formation and ( c and d ) c-di-GMP concentrations of PA14 cells cultured with terrein (50 μM) in the presence or absence of a different concentration of OdDHL or BHL for 9 h. Three independent experiments were performed in triplicate, and the mean ± SD values are displayed in each bar. * P
Figure Legend Snippet: Effects of exogenous QS ligands on the inhibition of biofilm and cellular c-di-GMP by terrein. ( a and b ) Biofilm formation and ( c and d ) c-di-GMP concentrations of PA14 cells cultured with terrein (50 μM) in the presence or absence of a different concentration of OdDHL or BHL for 9 h. Three independent experiments were performed in triplicate, and the mean ± SD values are displayed in each bar. * P

Techniques Used: Inhibition, Cell Culture, Concentration Assay

C-di-GMP levels in P . aeruginosa QS mutants. The cellular c-di-GMP concentration in biofilm cells of P . aeruginosa QS mutants ( lasI , lasR , rhlI , rhlR mutants) cultured for 9 h was compared with those in wild-type PA14. Three independent experiments were performed in triplicate, and the mean ± SD values are displayed in each bar. * P
Figure Legend Snippet: C-di-GMP levels in P . aeruginosa QS mutants. The cellular c-di-GMP concentration in biofilm cells of P . aeruginosa QS mutants ( lasI , lasR , rhlI , rhlR mutants) cultured for 9 h was compared with those in wild-type PA14. Three independent experiments were performed in triplicate, and the mean ± SD values are displayed in each bar. * P

Techniques Used: Concentration Assay, Cell Culture

Effects of terrein on cellular c-di-GMP. ( a and b ) Cellular c-di-GMP levels in biofilm cells of PAO1 cultured with different terrein or furanone C-30 concentrations for 9 h. After the biofilms were dissociated from the wells by gentle sonication, cellular c-di-GMP was extracted from the biofilm cells, measured, and normalized to the total proteins. ( c and d ) Comparison of biofilm formation by the P . aeruginosa wspF mutant versus wild-type PA14 cells cultured with different terrein or furanone C-30 concentrations for 9 h. Three independent experiments were performed in triplicate, and the mean ± SD values are displayed in each bar. * P
Figure Legend Snippet: Effects of terrein on cellular c-di-GMP. ( a and b ) Cellular c-di-GMP levels in biofilm cells of PAO1 cultured with different terrein or furanone C-30 concentrations for 9 h. After the biofilms were dissociated from the wells by gentle sonication, cellular c-di-GMP was extracted from the biofilm cells, measured, and normalized to the total proteins. ( c and d ) Comparison of biofilm formation by the P . aeruginosa wspF mutant versus wild-type PA14 cells cultured with different terrein or furanone C-30 concentrations for 9 h. Three independent experiments were performed in triplicate, and the mean ± SD values are displayed in each bar. * P

Techniques Used: Cell Culture, Sonication, Mutagenesis

Effects of terrein on virulence factor production and biofilm formation. ( a ) Chemical structures of terrein and furanone C-30. ( b ) Effects of terrein on virulence factor production and cell viability in P . aeruginosa PAO1. PAO1 cells were grown in LB medium containing various concentrations of terrein for 24 h, followed by the measurement of cell density at 600 nm, elastase activity, pyocyanin, and rhamnolipid in the culture supernatants. ( c ) Effects of terrein on biofilm formation and cell viability in P . aeruginosa . P . aeruginosa biofilms formed in the presence of terrein for 9 h. The biofilm cells attached to the well surface were assayed using crystal violet staining. ( d ) Effects of terrein on the antibiotic tolerance of biofilms. Preformed biofilms (6-h) of PAO1 cells were treated with ciprofloxacin (Cip) alone or together with DMSO or 300 μM terrein for 3 h. The biofilms were dissociated from the wells by gentle sonication, and the bacteria were enumerated by plating. Three independent experiments were performed in triplicate, and the mean ± standard deviation (SD) values are displayed in each bar. * P
Figure Legend Snippet: Effects of terrein on virulence factor production and biofilm formation. ( a ) Chemical structures of terrein and furanone C-30. ( b ) Effects of terrein on virulence factor production and cell viability in P . aeruginosa PAO1. PAO1 cells were grown in LB medium containing various concentrations of terrein for 24 h, followed by the measurement of cell density at 600 nm, elastase activity, pyocyanin, and rhamnolipid in the culture supernatants. ( c ) Effects of terrein on biofilm formation and cell viability in P . aeruginosa . P . aeruginosa biofilms formed in the presence of terrein for 9 h. The biofilm cells attached to the well surface were assayed using crystal violet staining. ( d ) Effects of terrein on the antibiotic tolerance of biofilms. Preformed biofilms (6-h) of PAO1 cells were treated with ciprofloxacin (Cip) alone or together with DMSO or 300 μM terrein for 3 h. The biofilms were dissociated from the wells by gentle sonication, and the bacteria were enumerated by plating. Three independent experiments were performed in triplicate, and the mean ± standard deviation (SD) values are displayed in each bar. * P

Techniques Used: Activity Assay, Staining, Sonication, Standard Deviation

35) Product Images from "Swarming motility and biofilm formation of Paenibacillus larvae, the etiological agent of American Foulbrood of honey bees (Apis mellifera)"

Article Title: Swarming motility and biofilm formation of Paenibacillus larvae, the etiological agent of American Foulbrood of honey bees (Apis mellifera)

Journal: Scientific Reports

doi: 10.1038/s41598-018-27193-8

Planktonic cells of P. larvae ERIC I (ATCC9545) and P. larvae ERIC II (DSM25430). Bacterial suspensions of P. larvae ERIC I (ATCC9545; A ) and P. larvae ERIC II (DSM25430; B ) in brain heart infusion (BHI) broth were incubated under constant agitation to prevent biofilm formation. Cultures of planktonic cells were stained with Congo red ( A , B ) while BHI medium stained with Congo red ( C ) or without staining ( D ) served as negative controls. Representative pictures are shown.
Figure Legend Snippet: Planktonic cells of P. larvae ERIC I (ATCC9545) and P. larvae ERIC II (DSM25430). Bacterial suspensions of P. larvae ERIC I (ATCC9545; A ) and P. larvae ERIC II (DSM25430; B ) in brain heart infusion (BHI) broth were incubated under constant agitation to prevent biofilm formation. Cultures of planktonic cells were stained with Congo red ( A , B ) while BHI medium stained with Congo red ( C ) or without staining ( D ) served as negative controls. Representative pictures are shown.

Techniques Used: Incubation, Staining

Involvement of paenilarvin in biofilm formation of P. larvae ERIC II. Wild-type P. larvae ERIC II (DSM25430 wt; A , C ) and a corresponding inactivation mutant for the paenilarvin gene cluster (DSM25430 Δ itu ; B , D ) were tested in biofilm assays. Bacterial suspensions in brain heart infusion (BHI) broth were incubated without agitation in six-well-plates at 37 °C for five ( A , B ) and six ( C , D ) days. For both strains, three biological replicates were performed both for obtaining pictures under unstained conditions ( A , B ) and after Congo red staining ( C , D ). Representative pictures are shown. ( E ) Biofilm formation was quantified via determining the amount of Conge red dye retained in the biofilms upon centrifugation. Data represent mean values ± SD of three independent experiments. The difference between the wild-type and corresponding inactivation mutant was not significant (p = 0.9889, Student’s t-test).
Figure Legend Snippet: Involvement of paenilarvin in biofilm formation of P. larvae ERIC II. Wild-type P. larvae ERIC II (DSM25430 wt; A , C ) and a corresponding inactivation mutant for the paenilarvin gene cluster (DSM25430 Δ itu ; B , D ) were tested in biofilm assays. Bacterial suspensions in brain heart infusion (BHI) broth were incubated without agitation in six-well-plates at 37 °C for five ( A , B ) and six ( C , D ) days. For both strains, three biological replicates were performed both for obtaining pictures under unstained conditions ( A , B ) and after Congo red staining ( C , D ). Representative pictures are shown. ( E ) Biofilm formation was quantified via determining the amount of Conge red dye retained in the biofilms upon centrifugation. Data represent mean values ± SD of three independent experiments. The difference between the wild-type and corresponding inactivation mutant was not significant (p = 0.9889, Student’s t-test).

Techniques Used: Mutagenesis, Incubation, Staining, Centrifugation

Biofilm formation of P. larvae ERIC I (ATCC9545) and P. larvae ERIC II (DSM25430) cultivated in static liquid. Bacterial suspensions of P. larvae ERIC I (ATCC9545; A , D , G ) and P. larvae ERIC II (DSM25430; B , E , H ) in brain heart infusion (BHI) broth were incubated without agitation in six-well-plates at 37 °C for five ( A , B ) and six ( D , E ) days, or in 96-well-plates for six days ( G , H ). For each P. larvae genotype, three biological replicates were performed both for obtaining pictures under unstained conditions ( A , B ), after Congo red staining of the extracellular matrix ( D , E ), and Crystal violet staining of the bacterial cells adherent to the well walls ( G , H ). Negative controls for each assay are shown in ( C , F , and I ). Representative pictures are shown.
Figure Legend Snippet: Biofilm formation of P. larvae ERIC I (ATCC9545) and P. larvae ERIC II (DSM25430) cultivated in static liquid. Bacterial suspensions of P. larvae ERIC I (ATCC9545; A , D , G ) and P. larvae ERIC II (DSM25430; B , E , H ) in brain heart infusion (BHI) broth were incubated without agitation in six-well-plates at 37 °C for five ( A , B ) and six ( D , E ) days, or in 96-well-plates for six days ( G , H ). For each P. larvae genotype, three biological replicates were performed both for obtaining pictures under unstained conditions ( A , B ), after Congo red staining of the extracellular matrix ( D , E ), and Crystal violet staining of the bacterial cells adherent to the well walls ( G , H ). Negative controls for each assay are shown in ( C , F , and I ). Representative pictures are shown.

Techniques Used: Incubation, Staining

Fluorescence microscopy of P. larvae floating biofilms. Bacterial suspensions of P. larvae ERIC I (ATCC9545; A , C ) and P. larvae ERIC II (DSM25430; B , D ) in Sf-900 II SFM medium supplemented with 30 µg/ml thioflavin S were incubated without agitation in 96-well-plates at 37 °C for six days. The thioflavin S-stained extracellular matrix in the floating biofilms was visualized using fluorescence microscopy; Z-stack processing was performed to obtain three-dimensional images of the wells containing the biofilms ( A , B ) and the region within the wells where the biofilms were located ( C , D ). Bars represent 20 µm.
Figure Legend Snippet: Fluorescence microscopy of P. larvae floating biofilms. Bacterial suspensions of P. larvae ERIC I (ATCC9545; A , C ) and P. larvae ERIC II (DSM25430; B , D ) in Sf-900 II SFM medium supplemented with 30 µg/ml thioflavin S were incubated without agitation in 96-well-plates at 37 °C for six days. The thioflavin S-stained extracellular matrix in the floating biofilms was visualized using fluorescence microscopy; Z-stack processing was performed to obtain three-dimensional images of the wells containing the biofilms ( A , B ) and the region within the wells where the biofilms were located ( C , D ). Bars represent 20 µm.

Techniques Used: Fluorescence, Microscopy, Incubation, Staining

36) Product Images from "TetR Family Regulator brpT Modulates Biofilm Formation in Streptococcus sanguinis"

Article Title: TetR Family Regulator brpT Modulates Biofilm Formation in Streptococcus sanguinis

Journal: PLoS ONE

doi: 10.1371/journal.pone.0169301

Deletion of gtfP in S . sanguinis decreases biofilm attachment and glucan synthesis. Wild-type, the gtfP mutant, Δ gtfP , and the double mutant, Δ brpT /Δ gtfP , were cultured in BM with 1% sucrose for 24 h anaerobically and analyzed. (A) Weak attachment of the Δ gtfP and the Δ brpT /Δ gtfP biofilm (pellicle) to the polystyrene surface and reduced biofilm biomass determined by CV staining. (B) Quantification of biofilm formation (OD 600 ). Quantification of (C) water soluble glucans, WSG and (D) water insoluble glucans, WIG accumulated within the biofilm. **, indicates significance with P
Figure Legend Snippet: Deletion of gtfP in S . sanguinis decreases biofilm attachment and glucan synthesis. Wild-type, the gtfP mutant, Δ gtfP , and the double mutant, Δ brpT /Δ gtfP , were cultured in BM with 1% sucrose for 24 h anaerobically and analyzed. (A) Weak attachment of the Δ gtfP and the Δ brpT /Δ gtfP biofilm (pellicle) to the polystyrene surface and reduced biofilm biomass determined by CV staining. (B) Quantification of biofilm formation (OD 600 ). Quantification of (C) water soluble glucans, WSG and (D) water insoluble glucans, WIG accumulated within the biofilm. **, indicates significance with P

Techniques Used: Mutagenesis, Cell Culture, Staining

SEM analysis further reveals altered biofilm morphology and an increase in filamentous structures. Biofilms formed by the wild-type SK36, the brpT mutant, Δ brpT , and the complemented mutant, Δ brpT_C , scanned under (A) 1000x magnification and (B) 20,000x magnification revealed an altered morphology and an increase in filamentous structures for Δ brpT compared to the wild-type and complemented mutant. White arrows indicate filamentous substances.
Figure Legend Snippet: SEM analysis further reveals altered biofilm morphology and an increase in filamentous structures. Biofilms formed by the wild-type SK36, the brpT mutant, Δ brpT , and the complemented mutant, Δ brpT_C , scanned under (A) 1000x magnification and (B) 20,000x magnification revealed an altered morphology and an increase in filamentous structures for Δ brpT compared to the wild-type and complemented mutant. White arrows indicate filamentous substances.

Techniques Used: Mutagenesis

Efficiency of glucan accumulation in S . sanguinis biofilms. S . sanguinis wild-type SK36, the brpT mutant Δ brpT and the complemented mutant Δ brpT _C were grown anaerobically for 24 h in BM medium containing 1% sucrose at 37°C. The amounts of (A) water soluble glucans and (B) water insoluble glucans in the biofilms were quantified using the phenol-sulfuric acid method and normalized to the concentration of genomic DNA.
Figure Legend Snippet: Efficiency of glucan accumulation in S . sanguinis biofilms. S . sanguinis wild-type SK36, the brpT mutant Δ brpT and the complemented mutant Δ brpT _C were grown anaerobically for 24 h in BM medium containing 1% sucrose at 37°C. The amounts of (A) water soluble glucans and (B) water insoluble glucans in the biofilms were quantified using the phenol-sulfuric acid method and normalized to the concentration of genomic DNA.

Techniques Used: Mutagenesis, Concentration Assay

Deletion of brpT alters the biofilm structure. (A) Wild-type S . sanguinis , SK36, the brpT mutant, Δ brpT , and the complemented mutant, Δ brpT _C were grown in BM as described in Materials and Methods . After 24-h growth, the biofilms were washed and stained with SYTO 9, and z -stacks of each were acquired by CLSM with a Plan-Neofluar ×10/0.3 objective lens. Representative orthogonal views from three independent experiments are displayed. (B) Quantification of biofilm thickness by CLSM for the wild-type, Δ brpT and Δ brpT _C. (C) Quantification of biofilm roughness for the wild-type, Δ brpT and Δ brpT _C. **, indicates significance with P
Figure Legend Snippet: Deletion of brpT alters the biofilm structure. (A) Wild-type S . sanguinis , SK36, the brpT mutant, Δ brpT , and the complemented mutant, Δ brpT _C were grown in BM as described in Materials and Methods . After 24-h growth, the biofilms were washed and stained with SYTO 9, and z -stacks of each were acquired by CLSM with a Plan-Neofluar ×10/0.3 objective lens. Representative orthogonal views from three independent experiments are displayed. (B) Quantification of biofilm thickness by CLSM for the wild-type, Δ brpT and Δ brpT _C. (C) Quantification of biofilm roughness for the wild-type, Δ brpT and Δ brpT _C. **, indicates significance with P

Techniques Used: Mutagenesis, Staining, Confocal Laser Scanning Microscopy

Deletion of brpT increases biofilm formation. S . sanguinis was cultured in BM supplemented with 1% sucrose, and biofilm biomass was determined by CV staining. Data from three biological replicates were averaged and the statistical significance between the brpT mutant, Δ brpT and the wild-type, SK36 or the complemented mutant, Δ brpT _C was determined by Student’s t -test. **, indicates significance with P
Figure Legend Snippet: Deletion of brpT increases biofilm formation. S . sanguinis was cultured in BM supplemented with 1% sucrose, and biofilm biomass was determined by CV staining. Data from three biological replicates were averaged and the statistical significance between the brpT mutant, Δ brpT and the wild-type, SK36 or the complemented mutant, Δ brpT _C was determined by Student’s t -test. **, indicates significance with P

Techniques Used: Cell Culture, Staining, Mutagenesis

37) Product Images from "Gentamicin promotes Staphylococcus aureus biofilms on silk suture 1"

Article Title: Gentamicin promotes Staphylococcus aureus biofilms on silk suture 1

Journal: The Journal of surgical research

doi: 10.1016/j.jss.2011.06.011

Biomass of biofilms formed by incubating silk suture with S. aureus RN6390 or ATCC 25923 for 24 hr, followed by overnight incubation with varying concentrations of gentamicin, with biomass measured (n≥5) as absorbance of crystal violet (A) and
Figure Legend Snippet: Biomass of biofilms formed by incubating silk suture with S. aureus RN6390 or ATCC 25923 for 24 hr, followed by overnight incubation with varying concentrations of gentamicin, with biomass measured (n≥5) as absorbance of crystal violet (A) and

Techniques Used: Incubation

Gentamicin susceptibility of planktonic and biofilm-associated S. aureus
Figure Legend Snippet: Gentamicin susceptibility of planktonic and biofilm-associated S. aureus

Techniques Used:

SEMs of biofilms formed by incubating silk suture with S. aureus ATCC 25923 for 24 hr, followed by overnight incubation with varying concentrations of gentamicin. A and B, Control biofilms after incubation with 0 µg/ml gentamicin, showing clusters
Figure Legend Snippet: SEMs of biofilms formed by incubating silk suture with S. aureus ATCC 25923 for 24 hr, followed by overnight incubation with varying concentrations of gentamicin. A and B, Control biofilms after incubation with 0 µg/ml gentamicin, showing clusters

Techniques Used: Incubation

Viability (A) and metabolic capacity (B) of intact biofilms formed by incubating silk suture with S. aureus RN6390 or ATCC 25923 for 24 hr, followed by overnight incubation with varying concentrations of gentamicin. A, Viable CFUs from sonicated biofilms
Figure Legend Snippet: Viability (A) and metabolic capacity (B) of intact biofilms formed by incubating silk suture with S. aureus RN6390 or ATCC 25923 for 24 hr, followed by overnight incubation with varying concentrations of gentamicin. A, Viable CFUs from sonicated biofilms

Techniques Used: Incubation, Sonication

38) Product Images from "The possible molecular mechanisms of farnesol on the antifungal resistance of C. albicans biofilms: the regulation of CYR1 and PDE2"

Article Title: The possible molecular mechanisms of farnesol on the antifungal resistance of C. albicans biofilms: the regulation of CYR1 and PDE2

Journal: BMC Microbiology

doi: 10.1186/s12866-018-1344-z

The expression of gene CYR1 in the presence of farnesol. Farnesol decreased the expression of CYR1 in the biofilms of the CAI4-pCaEXP strain at 12 and 24 h biofilm phases and decreased the expression of CYR1 in the biofilms of the CYR1OE strain at 6, 12, 24 and 36 h biofilm phases compared to the respective untreated controls. One-way analysis of variance (ANOVA) was employed to assess the statistical significance of differences in matched groups, while paired t-tests were performed for intra-group comparisons. *: p
Figure Legend Snippet: The expression of gene CYR1 in the presence of farnesol. Farnesol decreased the expression of CYR1 in the biofilms of the CAI4-pCaEXP strain at 12 and 24 h biofilm phases and decreased the expression of CYR1 in the biofilms of the CYR1OE strain at 6, 12, 24 and 36 h biofilm phases compared to the respective untreated controls. One-way analysis of variance (ANOVA) was employed to assess the statistical significance of differences in matched groups, while paired t-tests were performed for intra-group comparisons. *: p

Techniques Used: Expressing

The expression of PDE2 protein in C. albicans biofilms. a: WT: Farnesol untreated wild strain (CAI4-pCaEXP); WT + F: Farnesol treated wild strain (CAI4-pCaEXP); P: Farnesol untreated PDE2 -overexpressing strain (PDE2OE); P + F: Farnesol treated PDE2 -overexpressing strain (PDE2OE). a and b: Farnesol increased the expression of PDE2 of the CAI4-pCaEXP strain at 12, 24 and 36 h biofilm phases and increased the expression of PDE2 of the PDE2OE strain at 6, 12, 24 and 36 h biofilm phases compared to the respective untreated controls. Shown are means ± standard deviation of three independent experiments performed in duplicate. One-way analysis of variance (ANOVA) was employed to assess the statistical significance of differences in matched groups, while paired t-tests were performed for intra-group comparisons. *: p
Figure Legend Snippet: The expression of PDE2 protein in C. albicans biofilms. a: WT: Farnesol untreated wild strain (CAI4-pCaEXP); WT + F: Farnesol treated wild strain (CAI4-pCaEXP); P: Farnesol untreated PDE2 -overexpressing strain (PDE2OE); P + F: Farnesol treated PDE2 -overexpressing strain (PDE2OE). a and b: Farnesol increased the expression of PDE2 of the CAI4-pCaEXP strain at 12, 24 and 36 h biofilm phases and increased the expression of PDE2 of the PDE2OE strain at 6, 12, 24 and 36 h biofilm phases compared to the respective untreated controls. Shown are means ± standard deviation of three independent experiments performed in duplicate. One-way analysis of variance (ANOVA) was employed to assess the statistical significance of differences in matched groups, while paired t-tests were performed for intra-group comparisons. *: p

Techniques Used: Expressing, Standard Deviation

The expression of CYR1 protein in C. albicans biofilms. A: WT: Farnesol untreated wild strain (CAI4-pCaEXP); WT + F: Farnesol treated wild strain (CAI4-pCaEXP); C: Farnesol untreated CYR1 -overexpressing strain (CYR1OE); C + F: Farnesol treated CYR1 -overexpressing strain (CYR1OE). a and b: Farnesol decreased the expression of CYR1 of the CAI4-pCaEXP strain at 12, 24 and 36 h biofilm phases and decreased the expression of CYR1 of the CYR1OE strain at 6, 12 and 24 h biofilm phases compared to the respective untreated controls. Shown are means ± standard deviation of three independent experiments performed in duplicate. One-way analysis of variance (ANOVA) was employed to assess the statistical significance of differences in matched groups, while paired t-tests were performed for intra-group comparisons. *: p
Figure Legend Snippet: The expression of CYR1 protein in C. albicans biofilms. A: WT: Farnesol untreated wild strain (CAI4-pCaEXP); WT + F: Farnesol treated wild strain (CAI4-pCaEXP); C: Farnesol untreated CYR1 -overexpressing strain (CYR1OE); C + F: Farnesol treated CYR1 -overexpressing strain (CYR1OE). a and b: Farnesol decreased the expression of CYR1 of the CAI4-pCaEXP strain at 12, 24 and 36 h biofilm phases and decreased the expression of CYR1 of the CYR1OE strain at 6, 12 and 24 h biofilm phases compared to the respective untreated controls. Shown are means ± standard deviation of three independent experiments performed in duplicate. One-way analysis of variance (ANOVA) was employed to assess the statistical significance of differences in matched groups, while paired t-tests were performed for intra-group comparisons. *: p

Techniques Used: Expressing, Standard Deviation

The expression of gene PDE2 in the presence of farnesol. Farnesol increased the expression of PDE2 in the biofilms of the CAI4-pCaEXP strain at 12, 24 and 36 h biofilm phases and increased the expression of PDE2 in the biofilms of the PDE2OE strain at 6, 12, 24 and 36 h biofilm phases compared to the respective untreated controls. One-way analysis of variance (ANOVA) was employed to assess the statistical significance of differences in matched groups, while paired t-tests were performed for intra-group comparisons. *: p
Figure Legend Snippet: The expression of gene PDE2 in the presence of farnesol. Farnesol increased the expression of PDE2 in the biofilms of the CAI4-pCaEXP strain at 12, 24 and 36 h biofilm phases and increased the expression of PDE2 in the biofilms of the PDE2OE strain at 6, 12, 24 and 36 h biofilm phases compared to the respective untreated controls. One-way analysis of variance (ANOVA) was employed to assess the statistical significance of differences in matched groups, while paired t-tests were performed for intra-group comparisons. *: p

Techniques Used: Expressing

Confocal Laser Scanning Microscopy (CLSM) images of C. albicans biofilms formed from the CYR1OE and PDE2OE strains. A: a1–4: Farnesol untreated wild strain. b1–4: Farnesol untreated CYR1OE strain. c1–4: Farnesol untreated PDE2OE strain. d1–4: Farnesol treated wild strain. e1–4: Farnesol treated CYR1OE strain. f1–4: Farnesol treated PDE2OE strain. Scale bars represent 20 μm for 400× magnifications. The biofilms of d1–4, e1–4 and f1–4 consisted of fewer hyphae, but more pseudohyphae and yeast than did those of a1–4, b1–4 and c1–4. Additionally, the biofilms of b1–4 and c1–4 consisted of more hyphae and pseudohyphae than did that of a1–4. B: The biofilms exposed to farnesol were thinner than that without farnesol and the biofilms of the CYR1OE and PDE2OE strains were much thicker than that of the wild strain. Significance was calculated using one-way ANOVA with post ad-hoc Dunnett’s multiple comparison test. *: p
Figure Legend Snippet: Confocal Laser Scanning Microscopy (CLSM) images of C. albicans biofilms formed from the CYR1OE and PDE2OE strains. A: a1–4: Farnesol untreated wild strain. b1–4: Farnesol untreated CYR1OE strain. c1–4: Farnesol untreated PDE2OE strain. d1–4: Farnesol treated wild strain. e1–4: Farnesol treated CYR1OE strain. f1–4: Farnesol treated PDE2OE strain. Scale bars represent 20 μm for 400× magnifications. The biofilms of d1–4, e1–4 and f1–4 consisted of fewer hyphae, but more pseudohyphae and yeast than did those of a1–4, b1–4 and c1–4. Additionally, the biofilms of b1–4 and c1–4 consisted of more hyphae and pseudohyphae than did that of a1–4. B: The biofilms exposed to farnesol were thinner than that without farnesol and the biofilms of the CYR1OE and PDE2OE strains were much thicker than that of the wild strain. Significance was calculated using one-way ANOVA with post ad-hoc Dunnett’s multiple comparison test. *: p

Techniques Used: Confocal Laser Scanning Microscopy

The level of cAMP from C. albicans biofilms in the presence of farnesol. Farnesol decreased the intracellular cAMP levels in the biofilms of the CYR1OE strain, the PDE2OE strain, and the wild strain compared to those of the respective untreated controls. In addition, compared to the cAMP levels in the wild strain, the cAMP levels in the CYR1OE strain were significantly higher in all studied phases. One-way analysis of variance (ANOVA) was employed to assess the statistical significance of differences in matched groups, while paired t-tests were performed for intra-group comparisons. *: p
Figure Legend Snippet: The level of cAMP from C. albicans biofilms in the presence of farnesol. Farnesol decreased the intracellular cAMP levels in the biofilms of the CYR1OE strain, the PDE2OE strain, and the wild strain compared to those of the respective untreated controls. In addition, compared to the cAMP levels in the wild strain, the cAMP levels in the CYR1OE strain were significantly higher in all studied phases. One-way analysis of variance (ANOVA) was employed to assess the statistical significance of differences in matched groups, while paired t-tests were performed for intra-group comparisons. *: p

Techniques Used:

Scanning Electron Microscopy (SEM) images of C. albicans biofilms formed from the CYR1OE and PDE2 strains. A: a1–4: Farnesol untreated wild strain. b1–4: Farnesol untreated CYR1OE strain. c1–4: Farnesol untreated PDE2OE strain. d1–4: Farnesol treated wild strain. e1–4: Farnesol treated CYR1OE strain. f1–4: Farnesol treated PDE2OE strain. Magnification: 2000×. The surface of the cells in a1–4, d1–4 was uneven (red arrows), while the surface of cells in b1–4, c1–4, e1–4, f1–4 was smooth with spores (yellow arrows). The other results were the same as Fig. 2 A. B: The biofilms formed from the CYR1OE and PDE2OE strains exposed to farnesol consisted of shorter hyphae than that without farnesol. The hyphae extended much longer in the later phases (24 and 36 h) of the biofilms than did that in the early phases (6 h) . Significance was calculated using one-way ANOVA with post ad-hoc Dunnett’s multiple comparison test. *: p
Figure Legend Snippet: Scanning Electron Microscopy (SEM) images of C. albicans biofilms formed from the CYR1OE and PDE2 strains. A: a1–4: Farnesol untreated wild strain. b1–4: Farnesol untreated CYR1OE strain. c1–4: Farnesol untreated PDE2OE strain. d1–4: Farnesol treated wild strain. e1–4: Farnesol treated CYR1OE strain. f1–4: Farnesol treated PDE2OE strain. Magnification: 2000×. The surface of the cells in a1–4, d1–4 was uneven (red arrows), while the surface of cells in b1–4, c1–4, e1–4, f1–4 was smooth with spores (yellow arrows). The other results were the same as Fig. 2 A. B: The biofilms formed from the CYR1OE and PDE2OE strains exposed to farnesol consisted of shorter hyphae than that without farnesol. The hyphae extended much longer in the later phases (24 and 36 h) of the biofilms than did that in the early phases (6 h) . Significance was calculated using one-way ANOVA with post ad-hoc Dunnett’s multiple comparison test. *: p

Techniques Used: Electron Microscopy

39) Product Images from "The extracellular matrix protects Pseudomonas aeruginosa biofilms by limiting the penetration of tobramycin"

Article Title: The extracellular matrix protects Pseudomonas aeruginosa biofilms by limiting the penetration of tobramycin

Journal: Environmental microbiology

doi: 10.1111/1462-2920.12155

Cells at the periphery of the biofilm respond to tobramycin
Figure Legend Snippet: Cells at the periphery of the biofilm respond to tobramycin

Techniques Used:

Ability of the biofilm to limit penetration can be overcome with high concentrations of tobramycin
Figure Legend Snippet: Ability of the biofilm to limit penetration can be overcome with high concentrations of tobramycin

Techniques Used:

Metal cations facilitate the penetration of tobramycin into biofilms
Figure Legend Snippet: Metal cations facilitate the penetration of tobramycin into biofilms

Techniques Used:

Limiting penetration of tobramycin protects biofilm cells
Figure Legend Snippet: Limiting penetration of tobramycin protects biofilm cells

Techniques Used:

Biofilms limit the penetration of tobramycin into the biomass
Figure Legend Snippet: Biofilms limit the penetration of tobramycin into the biomass

Techniques Used:

40) Product Images from "Assessment of Antifungal Activity of Bakuchiol on Oral-Associated Candida spp."

Article Title: Assessment of Antifungal Activity of Bakuchiol on Oral-Associated Candida spp.

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

doi: 10.1155/2015/918624

Absorbance values of crystal violet solutions obtained from Candida biofilm formation following bakuchiol exposure. Data were represented as mean ± SD of three independent experiments performed in triplicate. Amphotericin B used as a positive control. Asterisk ( ∗ ) denotes the significant difference between treated samples and the untreated ones (one-way ANOVA, ∗ P
Figure Legend Snippet: Absorbance values of crystal violet solutions obtained from Candida biofilm formation following bakuchiol exposure. Data were represented as mean ± SD of three independent experiments performed in triplicate. Amphotericin B used as a positive control. Asterisk ( ∗ ) denotes the significant difference between treated samples and the untreated ones (one-way ANOVA, ∗ P

Techniques Used: Positive Control

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Incubation:

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Giemsa Stain:

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Inverted Microscopy:

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Staining:

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Sonication:

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    Fungal ITS region concentrations within drinking water <t>biofilms</t> developed under different chlorine concentrations. (A) The impact of chlorine concentration on gene concentration. For each regime, temporal changes are shown with respect to (B) Growth and (C) Re-growth phases. Data presented on a common plot were generated from the same qPCR run, p -values are from ANOVA analysis, n.s = no statistical significant difference, averages ( n = 5 or n = 4 see section “Bacterial and Fungal Quantification”) and standard deviation are presented.
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    Fungal ITS region concentrations within drinking water biofilms developed under different chlorine concentrations. (A) The impact of chlorine concentration on gene concentration. For each regime, temporal changes are shown with respect to (B) Growth and (C) Re-growth phases. Data presented on a common plot were generated from the same qPCR run, p -values are from ANOVA analysis, n.s = no statistical significant difference, averages ( n = 5 or n = 4 see section “Bacterial and Fungal Quantification”) and standard deviation are presented.

    Journal: Frontiers in Microbiology

    Article Title: Biofilm Microbiome (Re)Growth Dynamics in Drinking Water Distribution Systems Are Impacted by Chlorine Concentration

    doi: 10.3389/fmicb.2018.02519

    Figure Lengend Snippet: Fungal ITS region concentrations within drinking water biofilms developed under different chlorine concentrations. (A) The impact of chlorine concentration on gene concentration. For each regime, temporal changes are shown with respect to (B) Growth and (C) Re-growth phases. Data presented on a common plot were generated from the same qPCR run, p -values are from ANOVA analysis, n.s = no statistical significant difference, averages ( n = 5 or n = 4 see section “Bacterial and Fungal Quantification”) and standard deviation are presented.

    Article Snippet: Biofilm was removed from the outer coupons by standardized brushing into 30 mL sterile PBS, the biofilm suspensions were concentrated by filtering through a 47 mm diameter, 0.22 μm pore nitrocellulose membrane (Millipore, United States), as described in .

    Techniques: Concentration Assay, Generated, Real-time Polymerase Chain Reaction, Standard Deviation

    Resemblance between fungal community structures from biofilms developed under three different chlorine conditions and sampled at the time points indicated. Growth and Re-growth data are presented, global- R and p -values are from ANOSIM analysis for chlorine effects, gray lines indicate clusters of at least 30% similarity, based on group averages from hierarchical clustering; n.s. = no statistically significant difference.

    Journal: Frontiers in Microbiology

    Article Title: Biofilm Microbiome (Re)Growth Dynamics in Drinking Water Distribution Systems Are Impacted by Chlorine Concentration

    doi: 10.3389/fmicb.2018.02519

    Figure Lengend Snippet: Resemblance between fungal community structures from biofilms developed under three different chlorine conditions and sampled at the time points indicated. Growth and Re-growth data are presented, global- R and p -values are from ANOSIM analysis for chlorine effects, gray lines indicate clusters of at least 30% similarity, based on group averages from hierarchical clustering; n.s. = no statistically significant difference.

    Article Snippet: Biofilm was removed from the outer coupons by standardized brushing into 30 mL sterile PBS, the biofilm suspensions were concentrated by filtering through a 47 mm diameter, 0.22 μm pore nitrocellulose membrane (Millipore, United States), as described in .

    Techniques:

    Relative abundance of fungal classes (A) and genera (B) within biofilms from each chlorine regime, at each sample point. Classes in combination with the phyla they belong to (phylum_class) and genera are shown in the legend for each plot. “Unassigned” refers to sequences for which taxonomic information was unavailable, “Unknown” refers to sequences that were identified at a higher taxonomic level.

    Journal: Frontiers in Microbiology

    Article Title: Biofilm Microbiome (Re)Growth Dynamics in Drinking Water Distribution Systems Are Impacted by Chlorine Concentration

    doi: 10.3389/fmicb.2018.02519

    Figure Lengend Snippet: Relative abundance of fungal classes (A) and genera (B) within biofilms from each chlorine regime, at each sample point. Classes in combination with the phyla they belong to (phylum_class) and genera are shown in the legend for each plot. “Unassigned” refers to sequences for which taxonomic information was unavailable, “Unknown” refers to sequences that were identified at a higher taxonomic level.

    Article Snippet: Biofilm was removed from the outer coupons by standardized brushing into 30 mL sterile PBS, the biofilm suspensions were concentrated by filtering through a 47 mm diameter, 0.22 μm pore nitrocellulose membrane (Millipore, United States), as described in .

    Techniques:

    The number of shared and unique fungal mOTUs between biofilms from different chlorine concentrations throughout the (A) Growth and (B) Re-growth phases. Venn diagrams are based on presence/absence mOTU data.

    Journal: Frontiers in Microbiology

    Article Title: Biofilm Microbiome (Re)Growth Dynamics in Drinking Water Distribution Systems Are Impacted by Chlorine Concentration

    doi: 10.3389/fmicb.2018.02519

    Figure Lengend Snippet: The number of shared and unique fungal mOTUs between biofilms from different chlorine concentrations throughout the (A) Growth and (B) Re-growth phases. Venn diagrams are based on presence/absence mOTU data.

    Article Snippet: Biofilm was removed from the outer coupons by standardized brushing into 30 mL sterile PBS, the biofilm suspensions were concentrated by filtering through a 47 mm diameter, 0.22 μm pore nitrocellulose membrane (Millipore, United States), as described in .

    Techniques:

    Bacterial gene concentrations within drinking water biofilms developed under different chlorine concentrations. (A,B) show the impact of chlorine concentration, note the different y -axis scale in (B) . For each regime, temporal changes are shown with respect to (C) Growth and (D) Re-growth phases. Data presented on a common plot were generated from the same qPCR run, p -values are from ANOVA analysis, n.s = no statistical significant difference, averages ( n = 5) and standard deviation are presented.

    Journal: Frontiers in Microbiology

    Article Title: Biofilm Microbiome (Re)Growth Dynamics in Drinking Water Distribution Systems Are Impacted by Chlorine Concentration

    doi: 10.3389/fmicb.2018.02519

    Figure Lengend Snippet: Bacterial gene concentrations within drinking water biofilms developed under different chlorine concentrations. (A,B) show the impact of chlorine concentration, note the different y -axis scale in (B) . For each regime, temporal changes are shown with respect to (C) Growth and (D) Re-growth phases. Data presented on a common plot were generated from the same qPCR run, p -values are from ANOVA analysis, n.s = no statistical significant difference, averages ( n = 5) and standard deviation are presented.

    Article Snippet: Biofilm was removed from the outer coupons by standardized brushing into 30 mL sterile PBS, the biofilm suspensions were concentrated by filtering through a 47 mm diameter, 0.22 μm pore nitrocellulose membrane (Millipore, United States), as described in .

    Techniques: Concentration Assay, Generated, Real-time Polymerase Chain Reaction, Standard Deviation

    Resemblance between bacterial community structures from biofilms developed under three different chlorine conditions and sampled at the time points indicated. Growth and Re-growth data are presented, global- R and p -values are from ANOSIM analysis for chlorine effects. Full and dashed lines indicate clusters of at least 15 and 25% similarity, respectively, based on hierarchical clustering; n.s. = no statistically significant difference.

    Journal: Frontiers in Microbiology

    Article Title: Biofilm Microbiome (Re)Growth Dynamics in Drinking Water Distribution Systems Are Impacted by Chlorine Concentration

    doi: 10.3389/fmicb.2018.02519

    Figure Lengend Snippet: Resemblance between bacterial community structures from biofilms developed under three different chlorine conditions and sampled at the time points indicated. Growth and Re-growth data are presented, global- R and p -values are from ANOSIM analysis for chlorine effects. Full and dashed lines indicate clusters of at least 15 and 25% similarity, respectively, based on hierarchical clustering; n.s. = no statistically significant difference.

    Article Snippet: Biofilm was removed from the outer coupons by standardized brushing into 30 mL sterile PBS, the biofilm suspensions were concentrated by filtering through a 47 mm diameter, 0.22 μm pore nitrocellulose membrane (Millipore, United States), as described in .

    Techniques:

    Ecological indices of the bacterial communities within biofilms developed under different chlorine concentrations during (A) Growth and (B) Regrowth, averages ± standard deviation are presented.

    Journal: Frontiers in Microbiology

    Article Title: Biofilm Microbiome (Re)Growth Dynamics in Drinking Water Distribution Systems Are Impacted by Chlorine Concentration

    doi: 10.3389/fmicb.2018.02519

    Figure Lengend Snippet: Ecological indices of the bacterial communities within biofilms developed under different chlorine concentrations during (A) Growth and (B) Regrowth, averages ± standard deviation are presented.

    Article Snippet: Biofilm was removed from the outer coupons by standardized brushing into 30 mL sterile PBS, the biofilm suspensions were concentrated by filtering through a 47 mm diameter, 0.22 μm pore nitrocellulose membrane (Millipore, United States), as described in .

    Techniques: Standard Deviation

    The number of shared and unique bacterial mOTUs within and between biofilms from different chlorine concentrations throughout the Growth and Re-growth phases. Venn diagrams are based on presence/absence mOTU data.

    Journal: Frontiers in Microbiology

    Article Title: Biofilm Microbiome (Re)Growth Dynamics in Drinking Water Distribution Systems Are Impacted by Chlorine Concentration

    doi: 10.3389/fmicb.2018.02519

    Figure Lengend Snippet: The number of shared and unique bacterial mOTUs within and between biofilms from different chlorine concentrations throughout the Growth and Re-growth phases. Venn diagrams are based on presence/absence mOTU data.

    Article Snippet: Biofilm was removed from the outer coupons by standardized brushing into 30 mL sterile PBS, the biofilm suspensions were concentrated by filtering through a 47 mm diameter, 0.22 μm pore nitrocellulose membrane (Millipore, United States), as described in .

    Techniques:

    Drinking Water Distribution System Test Facility in which biofilms were developed. (A) The three independent tanks (0.49 m 3 ) and high density polyethylene (HDPE) loops of length 200 m and internal diameter of 79.3 mm, (B) The sections of each loop into which PWG coupons were inserted; M1–M3 indicate the location of the online chlorine meters, red circles highlight the location of flow meters used to control the hydraulics, and (C) Details of the PWG coupon dimensions. Figures are adapted from Fish et al. (2015 , 2018 ).

    Journal: Frontiers in Microbiology

    Article Title: Biofilm Microbiome (Re)Growth Dynamics in Drinking Water Distribution Systems Are Impacted by Chlorine Concentration

    doi: 10.3389/fmicb.2018.02519

    Figure Lengend Snippet: Drinking Water Distribution System Test Facility in which biofilms were developed. (A) The three independent tanks (0.49 m 3 ) and high density polyethylene (HDPE) loops of length 200 m and internal diameter of 79.3 mm, (B) The sections of each loop into which PWG coupons were inserted; M1–M3 indicate the location of the online chlorine meters, red circles highlight the location of flow meters used to control the hydraulics, and (C) Details of the PWG coupon dimensions. Figures are adapted from Fish et al. (2015 , 2018 ).

    Article Snippet: Biofilm was removed from the outer coupons by standardized brushing into 30 mL sterile PBS, the biofilm suspensions were concentrated by filtering through a 47 mm diameter, 0.22 μm pore nitrocellulose membrane (Millipore, United States), as described in .

    Techniques: Flow Cytometry, Fluorescence In Situ Hybridization

    Relative abundance of bacterial classes (A) and genera (B) within biofilms from each chlorine regime, at each sample point. Classes in combination with the phyla they belong to (phylum_class) and genera are shown in the legend for each plot. “Unassigned” refers to sequences for which taxonomic information was unavailable, “Unknown” refers to sequences that were identified at a higher taxonomic level. “Others” in (B) includes 55 genera which were present at

    Journal: Frontiers in Microbiology

    Article Title: Biofilm Microbiome (Re)Growth Dynamics in Drinking Water Distribution Systems Are Impacted by Chlorine Concentration

    doi: 10.3389/fmicb.2018.02519

    Figure Lengend Snippet: Relative abundance of bacterial classes (A) and genera (B) within biofilms from each chlorine regime, at each sample point. Classes in combination with the phyla they belong to (phylum_class) and genera are shown in the legend for each plot. “Unassigned” refers to sequences for which taxonomic information was unavailable, “Unknown” refers to sequences that were identified at a higher taxonomic level. “Others” in (B) includes 55 genera which were present at

    Article Snippet: Biofilm was removed from the outer coupons by standardized brushing into 30 mL sterile PBS, the biofilm suspensions were concentrated by filtering through a 47 mm diameter, 0.22 μm pore nitrocellulose membrane (Millipore, United States), as described in .

    Techniques:

    Ecological indices of the fungal communities within biofilms developed under different chlorine concentrations during (A) Growth and (B) Regrowth, averages ± standard deviations are presented.

    Journal: Frontiers in Microbiology

    Article Title: Biofilm Microbiome (Re)Growth Dynamics in Drinking Water Distribution Systems Are Impacted by Chlorine Concentration

    doi: 10.3389/fmicb.2018.02519

    Figure Lengend Snippet: Ecological indices of the fungal communities within biofilms developed under different chlorine concentrations during (A) Growth and (B) Regrowth, averages ± standard deviations are presented.

    Article Snippet: Biofilm was removed from the outer coupons by standardized brushing into 30 mL sterile PBS, the biofilm suspensions were concentrated by filtering through a 47 mm diameter, 0.22 μm pore nitrocellulose membrane (Millipore, United States), as described in .

    Techniques:

    Biofilm maturation and planktonic cell growth of Lactobacillus plantarum PA21. Cell viabilities of biofilm and planktonic Lactobacillus plantarum PA21 were measured over 7 days of biofilm development.

    Journal: Microbial Cell Factories

    Article Title: Inhibition of pathogenic and spoilage bacteria by a novel biofilm-forming Lactobacillus isolate: a potential host for the expression of heterologous proteins

    doi: 10.1186/s12934-015-0283-8

    Figure Lengend Snippet: Biofilm maturation and planktonic cell growth of Lactobacillus plantarum PA21. Cell viabilities of biofilm and planktonic Lactobacillus plantarum PA21 were measured over 7 days of biofilm development.

    Article Snippet: To observe planktonic cells, bacterial cells grown in suspensions were subjected to same methods as those in the biofilm before being transferred to an Isopore 0.2-μm membrane filter (Millipore, USA).

    Techniques:

    Comparative analysis of Lactobacillus plantarum PA21 in biofilm and planktonic culture. SEM analysis of a biofilm covered-surfaces, b biofilm cells, and c planktonic cells of A Lactobacillus plantarum ATCC 14917, B Lactobacillus plantarum PA21 in MRS broth after 24 h at 35°. Lactobacillus plantarum ATCC 14917 was used as positive control.

    Journal: Microbial Cell Factories

    Article Title: Inhibition of pathogenic and spoilage bacteria by a novel biofilm-forming Lactobacillus isolate: a potential host for the expression of heterologous proteins

    doi: 10.1186/s12934-015-0283-8

    Figure Lengend Snippet: Comparative analysis of Lactobacillus plantarum PA21 in biofilm and planktonic culture. SEM analysis of a biofilm covered-surfaces, b biofilm cells, and c planktonic cells of A Lactobacillus plantarum ATCC 14917, B Lactobacillus plantarum PA21 in MRS broth after 24 h at 35°. Lactobacillus plantarum ATCC 14917 was used as positive control.

    Article Snippet: To observe planktonic cells, bacterial cells grown in suspensions were subjected to same methods as those in the biofilm before being transferred to an Isopore 0.2-μm membrane filter (Millipore, USA).

    Techniques: Positive Control

    Biofilm formation on microtitre plates. Biofilm formation by two Lactobacillus strains on polystyrene microtitre plates following growth at 2 and 3 days at 30° and 35° in aerobic ( a ) and anaerobic ( b ) conditions in MRS broth: biofilms were stained with crystal violet, de-stained using 95% alcohol and the optical density at 595 nm of the alcoholic crystal violet solutions determined (OD optical density). Assays were performed three times for all 2 strains. Bars represent average values and standard errors of three observations.

    Journal: Microbial Cell Factories

    Article Title: Inhibition of pathogenic and spoilage bacteria by a novel biofilm-forming Lactobacillus isolate: a potential host for the expression of heterologous proteins

    doi: 10.1186/s12934-015-0283-8

    Figure Lengend Snippet: Biofilm formation on microtitre plates. Biofilm formation by two Lactobacillus strains on polystyrene microtitre plates following growth at 2 and 3 days at 30° and 35° in aerobic ( a ) and anaerobic ( b ) conditions in MRS broth: biofilms were stained with crystal violet, de-stained using 95% alcohol and the optical density at 595 nm of the alcoholic crystal violet solutions determined (OD optical density). Assays were performed three times for all 2 strains. Bars represent average values and standard errors of three observations.

    Article Snippet: To observe planktonic cells, bacterial cells grown in suspensions were subjected to same methods as those in the biofilm before being transferred to an Isopore 0.2-μm membrane filter (Millipore, USA).

    Techniques: Staining

    Number of viable Lactobacillus plantarum PA21 cells recovered from biofilm and planktonic cultures after contact with pathogens and food spoilage bacteria. Viable counts of Lb. plantarum PA21 biofilm cells ( a ), biofilm shed planktonic cells ( b ) and wild-type planktonic cells ( c ) were measured in co-cultures with Salmonella enterica, Pseudomonas fluorescens ATCC 13525, Aeromonas hydrophila ATCC 7965 and Bacillus cereus. Error bars indicate standard deviations of three independent experiments.

    Journal: Microbial Cell Factories

    Article Title: Inhibition of pathogenic and spoilage bacteria by a novel biofilm-forming Lactobacillus isolate: a potential host for the expression of heterologous proteins

    doi: 10.1186/s12934-015-0283-8

    Figure Lengend Snippet: Number of viable Lactobacillus plantarum PA21 cells recovered from biofilm and planktonic cultures after contact with pathogens and food spoilage bacteria. Viable counts of Lb. plantarum PA21 biofilm cells ( a ), biofilm shed planktonic cells ( b ) and wild-type planktonic cells ( c ) were measured in co-cultures with Salmonella enterica, Pseudomonas fluorescens ATCC 13525, Aeromonas hydrophila ATCC 7965 and Bacillus cereus. Error bars indicate standard deviations of three independent experiments.

    Article Snippet: To observe planktonic cells, bacterial cells grown in suspensions were subjected to same methods as those in the biofilm before being transferred to an Isopore 0.2-μm membrane filter (Millipore, USA).

    Techniques:

    Measure pH changes. pH values were measured during 6 days cultivation of Salmonella enterica, Pseudomonas fluorescens ATCC 13525, Aeromonas hydrophila ATCC 7965 and Bacillus cereus with Lb. plantarum PA21 biofilm.

    Journal: Microbial Cell Factories

    Article Title: Inhibition of pathogenic and spoilage bacteria by a novel biofilm-forming Lactobacillus isolate: a potential host for the expression of heterologous proteins

    doi: 10.1186/s12934-015-0283-8

    Figure Lengend Snippet: Measure pH changes. pH values were measured during 6 days cultivation of Salmonella enterica, Pseudomonas fluorescens ATCC 13525, Aeromonas hydrophila ATCC 7965 and Bacillus cereus with Lb. plantarum PA21 biofilm.

    Article Snippet: To observe planktonic cells, bacterial cells grown in suspensions were subjected to same methods as those in the biofilm before being transferred to an Isopore 0.2-μm membrane filter (Millipore, USA).

    Techniques:

    Efg1 regulates biofilm development by C. parapsilosis .A. The in vitro biofilm mass of smooth and concentric colonies from wild type (CLIB214), efg1 Δ / efg1 Δ deletion strains (including a mixture of strains LCP2, LCP5 and LCP8) and the reconstituted strain LCP7_RI was determined by measurement of dry weights. The average ± standard deviation from three independent measurements is shown. The panels below show biofilm growth on 24-well Nunc plates stained with crystal violet.B. In vivo biofilms were developed in the rat catheter model. Concentric wild-type (CLIB214) cells and smooth efg1 / efg1 deletion (LCP1) cells were allowed to develop for 24 h and then visualized by SEM at two magnifications.

    Journal: Molecular Microbiology

    Article Title: The APSES transcription factor Efg1 is a global regulator that controls morphogenesis and biofilm formation in Candida parapsilosis

    doi: 10.1111/mmi.12345

    Figure Lengend Snippet: Efg1 regulates biofilm development by C. parapsilosis .A. The in vitro biofilm mass of smooth and concentric colonies from wild type (CLIB214), efg1 Δ / efg1 Δ deletion strains (including a mixture of strains LCP2, LCP5 and LCP8) and the reconstituted strain LCP7_RI was determined by measurement of dry weights. The average ± standard deviation from three independent measurements is shown. The panels below show biofilm growth on 24-well Nunc plates stained with crystal violet.B. In vivo biofilms were developed in the rat catheter model. Concentric wild-type (CLIB214) cells and smooth efg1 / efg1 deletion (LCP1) cells were allowed to develop for 24 h and then visualized by SEM at two magnifications.

    Article Snippet: Biofilm from two wells was combined, filtered using a 0.8 μm Millipore filter, dried for 48 h at room temperature and weighed.

    Techniques: In Vitro, Standard Deviation, Staining, In Vivo

    a Schematic of experimental setup for the treatment of biofilm exposed to an e-scaffold with an illustration of electrochemical H 2 O 2 production. The electrodes are connected to a potentiostat (not shown in figure). Scientific Reports, 2015. 5, 14908. DOI: 10.1038/srep14908. © Creative Commons Attribution 4.0 International License. b Recovered biofilm cells treated with e-scaffold and tobramycin. c Treatment of biofilms and persister cells with e-scaffold

    Journal: NPJ Biofilms and Microbiomes

    Article Title: Eradication of Pseudomonas aeruginosa biofilms and persister cells using an electrochemical scaffold and enhanced antibiotic susceptibility

    doi: 10.1038/s41522-016-0003-0

    Figure Lengend Snippet: a Schematic of experimental setup for the treatment of biofilm exposed to an e-scaffold with an illustration of electrochemical H 2 O 2 production. The electrodes are connected to a potentiostat (not shown in figure). Scientific Reports, 2015. 5, 14908. DOI: 10.1038/srep14908. © Creative Commons Attribution 4.0 International License. b Recovered biofilm cells treated with e-scaffold and tobramycin. c Treatment of biofilms and persister cells with e-scaffold

    Article Snippet: Scanning electron microscopy For scanning electron microscopy (SEM) imaging, biofilms were grown for 24 h on UV-sterilized, 0.22-µm type GV membrane filters (Millipore, catalog ID #551200401) placed in sterile 6-well plates.

    Techniques:

    E-scaffold enhances tobramycin susceptibility in P. aeruginosa PAO1 biofilms. Bars represent means of logarithms of colony-forming units of viable biofilm cells. Error bars represent the standard error of the means from three biological replicates. The symbols *, **, ***, and **** represent significant differences in tobramycin susceptibility between e-scaffold treated biofilms + tobramycin and untreated biofilms + tobramycin ( n = 3, *, P = 0.002 ; **, ***, **** P ≤ 0.001; paired t -test)

    Journal: NPJ Biofilms and Microbiomes

    Article Title: Eradication of Pseudomonas aeruginosa biofilms and persister cells using an electrochemical scaffold and enhanced antibiotic susceptibility

    doi: 10.1038/s41522-016-0003-0

    Figure Lengend Snippet: E-scaffold enhances tobramycin susceptibility in P. aeruginosa PAO1 biofilms. Bars represent means of logarithms of colony-forming units of viable biofilm cells. Error bars represent the standard error of the means from three biological replicates. The symbols *, **, ***, and **** represent significant differences in tobramycin susceptibility between e-scaffold treated biofilms + tobramycin and untreated biofilms + tobramycin ( n = 3, *, P = 0.002 ; **, ***, **** P ≤ 0.001; paired t -test)

    Article Snippet: Scanning electron microscopy For scanning electron microscopy (SEM) imaging, biofilms were grown for 24 h on UV-sterilized, 0.22-µm type GV membrane filters (Millipore, catalog ID #551200401) placed in sterile 6-well plates.

    Techniques:

    Exposure to e-scaffold eradicates the persister cells in P. aeruginosa PAO1 biofilms. Bars represent means of logarithms of colony-forming units of viable biofilm cells. Error bars represent the standard errors of the means from three biological replicates. The symbol * denotes a significant difference compared to total biofilm cells ( n = 3, P

    Journal: NPJ Biofilms and Microbiomes

    Article Title: Eradication of Pseudomonas aeruginosa biofilms and persister cells using an electrochemical scaffold and enhanced antibiotic susceptibility

    doi: 10.1038/s41522-016-0003-0

    Figure Lengend Snippet: Exposure to e-scaffold eradicates the persister cells in P. aeruginosa PAO1 biofilms. Bars represent means of logarithms of colony-forming units of viable biofilm cells. Error bars represent the standard errors of the means from three biological replicates. The symbol * denotes a significant difference compared to total biofilm cells ( n = 3, P

    Article Snippet: Scanning electron microscopy For scanning electron microscopy (SEM) imaging, biofilms were grown for 24 h on UV-sterilized, 0.22-µm type GV membrane filters (Millipore, catalog ID #551200401) placed in sterile 6-well plates.

    Techniques:

    Scanning electron microscopy (SEM) images showing untreated, e-scaffold treated and exogenous H 2 O 2 treated P. aeruginosa PAO1 biofilm cells. Three representative images are shown for each treatment. Red arrows indicate cells showing a stressed membrane

    Journal: NPJ Biofilms and Microbiomes

    Article Title: Eradication of Pseudomonas aeruginosa biofilms and persister cells using an electrochemical scaffold and enhanced antibiotic susceptibility

    doi: 10.1038/s41522-016-0003-0

    Figure Lengend Snippet: Scanning electron microscopy (SEM) images showing untreated, e-scaffold treated and exogenous H 2 O 2 treated P. aeruginosa PAO1 biofilm cells. Three representative images are shown for each treatment. Red arrows indicate cells showing a stressed membrane

    Article Snippet: Scanning electron microscopy For scanning electron microscopy (SEM) imaging, biofilms were grown for 24 h on UV-sterilized, 0.22-µm type GV membrane filters (Millipore, catalog ID #551200401) placed in sterile 6-well plates.

    Techniques: Electron Microscopy

    E-scaffold increases OH• formation and membrane permeability. a Increase in fluorescence of HPF-stained e-scaffold treated and exogenous H 2 O 2 treated P. aeruginosa PAO1 biofilm cells indicates increased OH• formation compared to untreated biofilms. b Increase in fluorescence of propidium iodide (PI) indicates increased membrane permeability of P. aeruginosa PAO1 cells after exposure to e-scaffold. c Increase in fluorescence of 3, 3′-dipropylthiacarbocyanine iodide, a membrane potential sensitive dye, in e-scaffold treated P. aeruginosa PAO1 cells verifies bacterial membrane depolarization by e-scaffold resulting in increased membrane permeability. Error bars represent standard errors of means for at least three biological replicates. The symbol * indicates a significant difference from the untreated biofilm cells ( n = 3, P

    Journal: NPJ Biofilms and Microbiomes

    Article Title: Eradication of Pseudomonas aeruginosa biofilms and persister cells using an electrochemical scaffold and enhanced antibiotic susceptibility

    doi: 10.1038/s41522-016-0003-0

    Figure Lengend Snippet: E-scaffold increases OH• formation and membrane permeability. a Increase in fluorescence of HPF-stained e-scaffold treated and exogenous H 2 O 2 treated P. aeruginosa PAO1 biofilm cells indicates increased OH• formation compared to untreated biofilms. b Increase in fluorescence of propidium iodide (PI) indicates increased membrane permeability of P. aeruginosa PAO1 cells after exposure to e-scaffold. c Increase in fluorescence of 3, 3′-dipropylthiacarbocyanine iodide, a membrane potential sensitive dye, in e-scaffold treated P. aeruginosa PAO1 cells verifies bacterial membrane depolarization by e-scaffold resulting in increased membrane permeability. Error bars represent standard errors of means for at least three biological replicates. The symbol * indicates a significant difference from the untreated biofilm cells ( n = 3, P

    Article Snippet: Scanning electron microscopy For scanning electron microscopy (SEM) imaging, biofilms were grown for 24 h on UV-sterilized, 0.22-µm type GV membrane filters (Millipore, catalog ID #551200401) placed in sterile 6-well plates.

    Techniques: Permeability, Fluorescence, Staining