biofilm stain  (Thermo Fisher)


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    Structured Review

    Thermo Fisher biofilm stain
    Representative fluorescence microscopic images of implants (50x objective) after bacterial infection in vivo. The bar indicates 100 µ m. The red coloured areas in the picture correspond to the locations covered by <t>biofilms</t> of methicillin-resistant Staphylococcus aureus (MRSA). (a) HA coating at 1 day, (b) HA coating at 3 days, (c) HA coating at 7 days, (d) 3% Ag-HA coating at 1 day, (e) 3% Ag-HA coating at 3 days, and (f) 3% Ag-HA coating at 7 days. Calculated BCRs: (a) 16.0%, (b) 26.6%, (c) 30.1%, (d) 8.9%, (e) 14.6%, and (f) 13.7%.
    Biofilm Stain, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 90 stars, based on 1 article reviews
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    Images

    1) Product Images from "Silver-Containing Hydroxyapatite Coating Reduces Biofilm Formation by Methicillin-Resistant Staphylococcus aureus In Vitro and In Vivo"

    Article Title: Silver-Containing Hydroxyapatite Coating Reduces Biofilm Formation by Methicillin-Resistant Staphylococcus aureus In Vitro and In Vivo

    Journal: BioMed Research International

    doi: 10.1155/2016/8070597

    Representative fluorescence microscopic images of implants (50x objective) after bacterial infection in vivo. The bar indicates 100 µ m. The red coloured areas in the picture correspond to the locations covered by biofilms of methicillin-resistant Staphylococcus aureus (MRSA). (a) HA coating at 1 day, (b) HA coating at 3 days, (c) HA coating at 7 days, (d) 3% Ag-HA coating at 1 day, (e) 3% Ag-HA coating at 3 days, and (f) 3% Ag-HA coating at 7 days. Calculated BCRs: (a) 16.0%, (b) 26.6%, (c) 30.1%, (d) 8.9%, (e) 14.6%, and (f) 13.7%.
    Figure Legend Snippet: Representative fluorescence microscopic images of implants (50x objective) after bacterial infection in vivo. The bar indicates 100 µ m. The red coloured areas in the picture correspond to the locations covered by biofilms of methicillin-resistant Staphylococcus aureus (MRSA). (a) HA coating at 1 day, (b) HA coating at 3 days, (c) HA coating at 7 days, (d) 3% Ag-HA coating at 1 day, (e) 3% Ag-HA coating at 3 days, and (f) 3% Ag-HA coating at 7 days. Calculated BCRs: (a) 16.0%, (b) 26.6%, (c) 30.1%, (d) 8.9%, (e) 14.6%, and (f) 13.7%.

    Techniques Used: Fluorescence, Infection, In Vivo

    Box-and-whisker plots of biofilm coverage rates (BCRs) of implants after bacterial culture for 7 days and 14 days in vitro. In the 7-day experiment, 49 images of HA coating, including 14 images of 0.5% Ag-HA, 14 images of 1% Ag-HA, and 21 images of 3% Ag-HA, were used. In the 14-day experiment, 63 images of HA and 21 images of each group of Ag-HA were used. ∗ denotes significant differences between the BCRs of HA and each concentration of Ag-HA. The significance levels are as follows: 7 days: 0.5% Ag-HA, p = 0.011; 1% Ag-HA, p
    Figure Legend Snippet: Box-and-whisker plots of biofilm coverage rates (BCRs) of implants after bacterial culture for 7 days and 14 days in vitro. In the 7-day experiment, 49 images of HA coating, including 14 images of 0.5% Ag-HA, 14 images of 1% Ag-HA, and 21 images of 3% Ag-HA, were used. In the 14-day experiment, 63 images of HA and 21 images of each group of Ag-HA were used. ∗ denotes significant differences between the BCRs of HA and each concentration of Ag-HA. The significance levels are as follows: 7 days: 0.5% Ag-HA, p = 0.011; 1% Ag-HA, p

    Techniques Used: Whisker Assay, In Vitro, Concentration Assay

    Related Articles

    Staining:

    Article Title: Antibodies directed against Integration Host Factor Mediate Biofilm Clearance from Nasopore®
    Article Snippet: .. The remaining wells were stained with 50 uL of Filmtracer Biofilm stain (Life Technologies, Grand Island, NY). .. Filmtracer Biofilm stain was used in light of the fact that Live/Dead stain was taken up by Nasopore and thus resulted in an inability to subtract Nasopore mass when performing COMSTAT analysis between biofilms from confocal imaging.

    Article Title: Silver-Containing Hydroxyapatite Coating Reduces Biofilm Formation by Methicillin-Resistant Staphylococcus aureus In Vitro and In Vivo
    Article Snippet: .. Calculation of Biofilm Coverage Rates (BCRs) All disks (from in vitro and in vivo experiments) were rinsed twice with 500 μ L of sterile PBS and stained with biofilm stain (FilmTracer calcein red-orange biofilm stain, Thermo Fisher Scientific) for 1 h. After staining, disks were washed twice with 500 μ L of sterile PBS. .. Thereafter, biofilm formation on each disk was observed under a fluorescence microscope (Axioplan 2; ZEISS, Jena, Germany).

    Article Title: Host and Pathogen Hyaluronan Signal Through Human Siglec-9 to Suppress Neutrophil Activation
    Article Snippet: .. 5 × 105 neutrophils were labeled with FilmTracer Calcein green (Invitrogen) vital staining according to the manufacturer instructions. .. 1 µg/ml HMW-HA (Sigma) was bound to 96-well plate and blocked with 3% BSA, then neutrophils plated and incubated at 37°C for 30 min. After washing the plate with 1% BSA in Hanks’ balanced salt solution (HBSS), adherent cells were visualized under a fluorescent microscope and enumerated by counting in a hemocytometer.

    In Vivo:

    Article Title: Silver-Containing Hydroxyapatite Coating Reduces Biofilm Formation by Methicillin-Resistant Staphylococcus aureus In Vitro and In Vivo
    Article Snippet: .. Calculation of Biofilm Coverage Rates (BCRs) All disks (from in vitro and in vivo experiments) were rinsed twice with 500 μ L of sterile PBS and stained with biofilm stain (FilmTracer calcein red-orange biofilm stain, Thermo Fisher Scientific) for 1 h. After staining, disks were washed twice with 500 μ L of sterile PBS. .. Thereafter, biofilm formation on each disk was observed under a fluorescence microscope (Axioplan 2; ZEISS, Jena, Germany).

    Incubation:

    Article Title: MRP8/14 Enhances Corneal Susceptibility to Pseudomonas aeruginosa Infection by Amplifying Inflammatory Responses
    Article Snippet: .. In brief, PA was incubated with Filmtracer Green Biofilm (FTGB, 1:50; Invitrogen) at room temperature for 30 minutes protected from light, and then rinsed gently with sterilized water. .. RAW264.7 cells were transfected with siMRP8/14 or siCTL for 24 hours, and then challenged with FTGB-stained PA at a multiplicity of infection (MOI) of 25.

    Labeling:

    Article Title: Host and Pathogen Hyaluronan Signal Through Human Siglec-9 to Suppress Neutrophil Activation
    Article Snippet: .. 5 × 105 neutrophils were labeled with FilmTracer Calcein green (Invitrogen) vital staining according to the manufacturer instructions. .. 1 µg/ml HMW-HA (Sigma) was bound to 96-well plate and blocked with 3% BSA, then neutrophils plated and incubated at 37°C for 30 min. After washing the plate with 1% BSA in Hanks’ balanced salt solution (HBSS), adherent cells were visualized under a fluorescent microscope and enumerated by counting in a hemocytometer.

    In Vitro:

    Article Title: Silver-Containing Hydroxyapatite Coating Reduces Biofilm Formation by Methicillin-Resistant Staphylococcus aureus In Vitro and In Vivo
    Article Snippet: .. Calculation of Biofilm Coverage Rates (BCRs) All disks (from in vitro and in vivo experiments) were rinsed twice with 500 μ L of sterile PBS and stained with biofilm stain (FilmTracer calcein red-orange biofilm stain, Thermo Fisher Scientific) for 1 h. After staining, disks were washed twice with 500 μ L of sterile PBS. .. Thereafter, biofilm formation on each disk was observed under a fluorescence microscope (Axioplan 2; ZEISS, Jena, Germany).

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    Thermo Fisher filmtracertm sypro ruby biofilm matrix stain
    LIVE/DEAD® Baclight TM and FilmTracer TM <t>SYPRO®</t> Ruby <t>Biofilm</t> Matrix Stain of 48-h-old biofilms of P . aeruginosa PA01 (A) and S . aureus Xen 30 (B). For each strain, images were taken of biofilms that formed in (a) the absence of nanofibers and copper, (b) nanofibers without copper particles (CF) and (c) copper-containing nanofibers (Cu-F). The images in column 1 is after propidium iodide staining, column 2 after ruby staining and column 3 after SYTO® 9 staining. The image in column 4 is an overlap of all stains.
    Filmtracertm Sypro Ruby Biofilm Matrix Stain, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    Thermo Fisher biofilm matrix stain
    Exophiala dermatitidis (Ed) <t>biofilm</t> formed after 24 or 48 h at 36°C in artificial sputum medium (ASM). (A) Biofilms formed in mono- (Ed pure) and co-cultures with Pseudomonas aeruginosa (Pa). Biofilms were estimated by detachment of biofilm with 0.1% dithiothreitol and subsequent count of colony-forming units (CFU per mL). (B) Biofilms in Transwell permeable support. Ed suspension for biofilm formation was placed into the well, whereas Pa was placed in the insert. As a control, pure ASM was used. Unpaired t -test, * P
    Biofilm Matrix Stain, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    LIVE/DEAD® Baclight TM and FilmTracer TM SYPRO® Ruby Biofilm Matrix Stain of 48-h-old biofilms of P . aeruginosa PA01 (A) and S . aureus Xen 30 (B). For each strain, images were taken of biofilms that formed in (a) the absence of nanofibers and copper, (b) nanofibers without copper particles (CF) and (c) copper-containing nanofibers (Cu-F). The images in column 1 is after propidium iodide staining, column 2 after ruby staining and column 3 after SYTO® 9 staining. The image in column 4 is an overlap of all stains.

    Journal: PLoS ONE

    Article Title: Copper-Containing Anti-Biofilm Nanofiber Scaffolds as a Wound Dressing Material

    doi: 10.1371/journal.pone.0152755

    Figure Lengend Snippet: LIVE/DEAD® Baclight TM and FilmTracer TM SYPRO® Ruby Biofilm Matrix Stain of 48-h-old biofilms of P . aeruginosa PA01 (A) and S . aureus Xen 30 (B). For each strain, images were taken of biofilms that formed in (a) the absence of nanofibers and copper, (b) nanofibers without copper particles (CF) and (c) copper-containing nanofibers (Cu-F). The images in column 1 is after propidium iodide staining, column 2 after ruby staining and column 3 after SYTO® 9 staining. The image in column 4 is an overlap of all stains.

    Article Snippet: The LIVE/DEAD® BaclightTM Bacterial Viability kit and FilmTracerTM SYPRO® Ruby Biofilm Matrix Stain were from Thermo Fisher Scientific (Massachusetts, USA).

    Techniques: Staining

    Macromolecular composition of the S . epidermidis biofilm matrix. CLSM analysis of the biofilm matrix composition of S . epidermidis ST10002 (PIA + ) and AZ39 (PIA - ). WGA-OG488 staining shows polysaccharide content attributed to PIA production. SYPRO-Ruby staining shows proteins contained in the biofilm matrix and DAPI staining was used to detect DNA and visualize bacteria. TSBg biofilms were observed with a 40x objective. Scale bar = 20 μm. PC biofilms were observed with a 63x objective. Scale bar = 5 μm. TSBg coupons were stored at 4°C in PBS 1x before staining while PC coupons were rinsed with PBS 1X, dried, and stored at -20°C before staining.

    Journal: PLoS ONE

    Article Title: The peptidoglycan and biofilm matrix of Staphylococcus epidermidis undergo structural changes when exposed to human platelets

    doi: 10.1371/journal.pone.0211132

    Figure Lengend Snippet: Macromolecular composition of the S . epidermidis biofilm matrix. CLSM analysis of the biofilm matrix composition of S . epidermidis ST10002 (PIA + ) and AZ39 (PIA - ). WGA-OG488 staining shows polysaccharide content attributed to PIA production. SYPRO-Ruby staining shows proteins contained in the biofilm matrix and DAPI staining was used to detect DNA and visualize bacteria. TSBg biofilms were observed with a 40x objective. Scale bar = 20 μm. PC biofilms were observed with a 63x objective. Scale bar = 5 μm. TSBg coupons were stored at 4°C in PBS 1x before staining while PC coupons were rinsed with PBS 1X, dried, and stored at -20°C before staining.

    Article Snippet: Biofilms were then labelled with three fluorescent dyes as follows: (I) Wheat germ agglutinin (WGA) conjugated with Oregon Green-488 (Thermo Fisher Scientific, Waltham, MA, USA) (WGA-OG488), which labels polysaccharides by binding to N-acetylglucosamine and sialic acid residues, 5 μm in dH2 O/15 min. (II) Film tracer SYPRO Ruby Biofilm Matrix Stain (Thermo Fisher Scientific, Waltham, MA, USA), which labels most types of proteins, for 15 min prior to live confocal imaging (III) Coupons were then mounted on slides under coverslips in DAPI-Fluoromount-G (Electron Microscopy Sciences, Hatfield, PA, USA), containing 4’,6-diamidino-2-phenylindole (DAPI), which labels DNA.

    Techniques: Confocal Laser Scanning Microscopy, Whole Genome Amplification, Staining

    Induction of luxS expression promotes biofilm development in a stationary system. Stationary biofilms were established by H. influenzae strains NT Hi 86-028NP, NT Hi 86-028NP luxS , and WES204 in sBHI medium with or without 5 mM xylose, as indicated, and stained with a BacLight live/dead reagent (Molecular Probes) for confocal microscopy. (A to D) Vertical z-series images were compiled to generate representative volume views of 12-h biofilms formed by NT Hi 86-028NP (A), WES204 induced by xylose for 12 h (B), NT Hi 86-028NP luxS (C), and uninduced WES204 (D). (E to J) z-series images of biofilms formed for 12 h (E and F), 24 h (G and H), or 48 h (I and J) were exported into Matlab for COMSTAT analysis of biofilm average thickness (E, G, and I) and total biomass (F, H, and J). Graphs were derived from 10 individual fields of view per well in 4 independent biofilms per wells, and error bars represent means and standard deviations.

    Journal: Infection and Immunity

    Article Title: Autoinducer 2 (AI-2) Production by Nontypeable Haemophilus influenzae 86-028NP Promotes Expression of a Predicted Glycosyltransferase That Is a Determinant of Biofilm Maturation, Prevention of Dispersal, and Persistence In Vivo

    doi: 10.1128/IAI.00506-18

    Figure Lengend Snippet: Induction of luxS expression promotes biofilm development in a stationary system. Stationary biofilms were established by H. influenzae strains NT Hi 86-028NP, NT Hi 86-028NP luxS , and WES204 in sBHI medium with or without 5 mM xylose, as indicated, and stained with a BacLight live/dead reagent (Molecular Probes) for confocal microscopy. (A to D) Vertical z-series images were compiled to generate representative volume views of 12-h biofilms formed by NT Hi 86-028NP (A), WES204 induced by xylose for 12 h (B), NT Hi 86-028NP luxS (C), and uninduced WES204 (D). (E to J) z-series images of biofilms formed for 12 h (E and F), 24 h (G and H), or 48 h (I and J) were exported into Matlab for COMSTAT analysis of biofilm average thickness (E, G, and I) and total biomass (F, H, and J). Graphs were derived from 10 individual fields of view per well in 4 independent biofilms per wells, and error bars represent means and standard deviations.

    Article Snippet: In order to compare the biofilm matrices, comparable biofilm cultures of both strains were stained with FilmTracer SYPRO ruby red biofilm matrix stain (Thermo Fisher), and similar confocal Z-stack images of NT Hi 86-028NP and NT Hi 86-028NP gstA were generated and compared ( ).

    Techniques: Expressing, Staining, Confocal Microscopy, Derivative Assay

    Continuous luxS expression prevents NT Hi 86-028NP biofilm dispersal. (A) Biofilms were established by H. influenzae gfp -expressing strains NT Hi 86-028NP and WES204 for 96 h under continuous medium flow, with confocal microscopy every 24 h for COMSTAT analysis of average biofilm thickness. NT Hi 86-028NP biofilms were cultured in sBHI medium, and WES204 was cultured in sBHI medium supplemented with 5 mM xylose. Values were derived from 3 independent replicates per variable, and error bars represent means and standard deviations. **, P

    Journal: Infection and Immunity

    Article Title: Autoinducer 2 (AI-2) Production by Nontypeable Haemophilus influenzae 86-028NP Promotes Expression of a Predicted Glycosyltransferase That Is a Determinant of Biofilm Maturation, Prevention of Dispersal, and Persistence In Vivo

    doi: 10.1128/IAI.00506-18

    Figure Lengend Snippet: Continuous luxS expression prevents NT Hi 86-028NP biofilm dispersal. (A) Biofilms were established by H. influenzae gfp -expressing strains NT Hi 86-028NP and WES204 for 96 h under continuous medium flow, with confocal microscopy every 24 h for COMSTAT analysis of average biofilm thickness. NT Hi 86-028NP biofilms were cultured in sBHI medium, and WES204 was cultured in sBHI medium supplemented with 5 mM xylose. Values were derived from 3 independent replicates per variable, and error bars represent means and standard deviations. **, P

    Article Snippet: In order to compare the biofilm matrices, comparable biofilm cultures of both strains were stained with FilmTracer SYPRO ruby red biofilm matrix stain (Thermo Fisher), and similar confocal Z-stack images of NT Hi 86-028NP and NT Hi 86-028NP gstA were generated and compared ( ).

    Techniques: Expressing, Flow Cytometry, Confocal Microscopy, Cell Culture, Derivative Assay

    Modulation of luxS expression alters biofilm development and dispersal. Biofilms were established by gfp -expressing WES204 under continuous-medium-flow conditions in sBHI medium or sBHI medium supplemented with 5 mM xylose, as indicated, and imaged by confocal microscopy at 24, 27, and 48 h postinoculation for COMSTAT analysis of total biofilm biomass (A) and average thickness (B). At 24 h postinoculation, the medium source was switched, when indicated, from sBHI medium to sBHI medium supplemented with xylose or vice versa and maintained for the remainder of the experiment. Data were derived from 3 independent continuous-flow biofilms per variable, and error bars represent means and standard deviations.

    Journal: Infection and Immunity

    Article Title: Autoinducer 2 (AI-2) Production by Nontypeable Haemophilus influenzae 86-028NP Promotes Expression of a Predicted Glycosyltransferase That Is a Determinant of Biofilm Maturation, Prevention of Dispersal, and Persistence In Vivo

    doi: 10.1128/IAI.00506-18

    Figure Lengend Snippet: Modulation of luxS expression alters biofilm development and dispersal. Biofilms were established by gfp -expressing WES204 under continuous-medium-flow conditions in sBHI medium or sBHI medium supplemented with 5 mM xylose, as indicated, and imaged by confocal microscopy at 24, 27, and 48 h postinoculation for COMSTAT analysis of total biofilm biomass (A) and average thickness (B). At 24 h postinoculation, the medium source was switched, when indicated, from sBHI medium to sBHI medium supplemented with xylose or vice versa and maintained for the remainder of the experiment. Data were derived from 3 independent continuous-flow biofilms per variable, and error bars represent means and standard deviations.

    Article Snippet: In order to compare the biofilm matrices, comparable biofilm cultures of both strains were stained with FilmTracer SYPRO ruby red biofilm matrix stain (Thermo Fisher), and similar confocal Z-stack images of NT Hi 86-028NP and NT Hi 86-028NP gstA were generated and compared ( ).

    Techniques: Expressing, Flow Cytometry, Confocal Microscopy, Derivative Assay

    Transcription of luxS decreases as NT Hi 86-028NP biofilms approach dispersal. NT Hi 86-028NP stationary biofilms were established in sBHI medium for 6, 12, 24, 48, and 72 h for isolation of RNA and measurement of luxS transcripts by real-time RT-PCR. Data were normalized and are expressed as ratios of luxS to gyrA . Each measure represents data for 5 independent samples, and error bars represent means and standard deviations.

    Journal: Infection and Immunity

    Article Title: Autoinducer 2 (AI-2) Production by Nontypeable Haemophilus influenzae 86-028NP Promotes Expression of a Predicted Glycosyltransferase That Is a Determinant of Biofilm Maturation, Prevention of Dispersal, and Persistence In Vivo

    doi: 10.1128/IAI.00506-18

    Figure Lengend Snippet: Transcription of luxS decreases as NT Hi 86-028NP biofilms approach dispersal. NT Hi 86-028NP stationary biofilms were established in sBHI medium for 6, 12, 24, 48, and 72 h for isolation of RNA and measurement of luxS transcripts by real-time RT-PCR. Data were normalized and are expressed as ratios of luxS to gyrA . Each measure represents data for 5 independent samples, and error bars represent means and standard deviations.

    Article Snippet: In order to compare the biofilm matrices, comparable biofilm cultures of both strains were stained with FilmTracer SYPRO ruby red biofilm matrix stain (Thermo Fisher), and similar confocal Z-stack images of NT Hi 86-028NP and NT Hi 86-028NP gstA were generated and compared ( ).

    Techniques: Isolation, Quantitative RT-PCR

    gstA has a significant impact on NT Hi 86-028NP tissue-associated (biofilm) bacterial loads in experimental chinchilla otitis media infections. Chinchillas (6 per group) were experimentally infected via the transbullar route with ∼10 3 CFU/ear of NT Hi 86-028NP or NT Hi 86-028NP gstA (see Materials and Methods). Groups of animals were euthanized at 1 week, 2 weeks, and 3 weeks postinfection, and bacterial loads in effusion and bullar tissue homogenate samples were determined by plate counts. *, statistically significant differences between groups ( P

    Journal: Infection and Immunity

    Article Title: Autoinducer 2 (AI-2) Production by Nontypeable Haemophilus influenzae 86-028NP Promotes Expression of a Predicted Glycosyltransferase That Is a Determinant of Biofilm Maturation, Prevention of Dispersal, and Persistence In Vivo

    doi: 10.1128/IAI.00506-18

    Figure Lengend Snippet: gstA has a significant impact on NT Hi 86-028NP tissue-associated (biofilm) bacterial loads in experimental chinchilla otitis media infections. Chinchillas (6 per group) were experimentally infected via the transbullar route with ∼10 3 CFU/ear of NT Hi 86-028NP or NT Hi 86-028NP gstA (see Materials and Methods). Groups of animals were euthanized at 1 week, 2 weeks, and 3 weeks postinfection, and bacterial loads in effusion and bullar tissue homogenate samples were determined by plate counts. *, statistically significant differences between groups ( P

    Article Snippet: In order to compare the biofilm matrices, comparable biofilm cultures of both strains were stained with FilmTracer SYPRO ruby red biofilm matrix stain (Thermo Fisher), and similar confocal Z-stack images of NT Hi 86-028NP and NT Hi 86-028NP gstA were generated and compared ( ).

    Techniques: Infection

    Exophiala dermatitidis (Ed) biofilm formed after 24 or 48 h at 36°C in artificial sputum medium (ASM). (A) Biofilms formed in mono- (Ed pure) and co-cultures with Pseudomonas aeruginosa (Pa). Biofilms were estimated by detachment of biofilm with 0.1% dithiothreitol and subsequent count of colony-forming units (CFU per mL). (B) Biofilms in Transwell permeable support. Ed suspension for biofilm formation was placed into the well, whereas Pa was placed in the insert. As a control, pure ASM was used. Unpaired t -test, * P

    Journal: Frontiers in Microbiology

    Article Title: Phenotypical Characteristics of the Black Yeast Exophiala dermatitidis Are Affected by Pseudomonas aeruginosa in an Artificial Sputum Medium Mimicking Cystic Fibrosis–Like Conditions

    doi: 10.3389/fmicb.2020.00471

    Figure Lengend Snippet: Exophiala dermatitidis (Ed) biofilm formed after 24 or 48 h at 36°C in artificial sputum medium (ASM). (A) Biofilms formed in mono- (Ed pure) and co-cultures with Pseudomonas aeruginosa (Pa). Biofilms were estimated by detachment of biofilm with 0.1% dithiothreitol and subsequent count of colony-forming units (CFU per mL). (B) Biofilms in Transwell permeable support. Ed suspension for biofilm formation was placed into the well, whereas Pa was placed in the insert. As a control, pure ASM was used. Unpaired t -test, * P

    Article Snippet: ECM thickness was determined by staining with Invitrogen FilmTracer SYPRO ruby biofilm matrix stain (Thermo Fisher Scientific, Waltham, MA, United States) and subsequent CLSM.

    Techniques:

    Survival of Galleria mellonella after infection with Exophiala dermatitidis (Ed; Strain P2) with or without Pseudomonas aeruginosa (Pa) culture filtrate of strain PA07 (A/B), PA14 wild-type (wt; C/D), PA14 ΔlasR (E/F), and PA14 ΔrhlR (G/H). As a control, sterile phosphate-buffered saline (PBS) was used. (A, C, E, G) Planktonic culture filtrate was added to injection suspension (S). (B, D, F, H) Biofilm culture filtrate (BFS) 24 or 48 h old was added to the injection suspension. N = 30. Log-rank (Mantel-Cox) test for statistical significance: * P

    Journal: Frontiers in Microbiology

    Article Title: Phenotypical Characteristics of the Black Yeast Exophiala dermatitidis Are Affected by Pseudomonas aeruginosa in an Artificial Sputum Medium Mimicking Cystic Fibrosis–Like Conditions

    doi: 10.3389/fmicb.2020.00471

    Figure Lengend Snippet: Survival of Galleria mellonella after infection with Exophiala dermatitidis (Ed; Strain P2) with or without Pseudomonas aeruginosa (Pa) culture filtrate of strain PA07 (A/B), PA14 wild-type (wt; C/D), PA14 ΔlasR (E/F), and PA14 ΔrhlR (G/H). As a control, sterile phosphate-buffered saline (PBS) was used. (A, C, E, G) Planktonic culture filtrate was added to injection suspension (S). (B, D, F, H) Biofilm culture filtrate (BFS) 24 or 48 h old was added to the injection suspension. N = 30. Log-rank (Mantel-Cox) test for statistical significance: * P

    Article Snippet: ECM thickness was determined by staining with Invitrogen FilmTracer SYPRO ruby biofilm matrix stain (Thermo Fisher Scientific, Waltham, MA, United States) and subsequent CLSM.

    Techniques: Infection, Injection

    Extracellular matrix (ECM) thickness (μm) of Exophiala dermatitidis mono- (Ed pure) and co-culture biofilms with Pseudomonas aeruginosa (Pa). N = 10.

    Journal: Frontiers in Microbiology

    Article Title: Phenotypical Characteristics of the Black Yeast Exophiala dermatitidis Are Affected by Pseudomonas aeruginosa in an Artificial Sputum Medium Mimicking Cystic Fibrosis–Like Conditions

    doi: 10.3389/fmicb.2020.00471

    Figure Lengend Snippet: Extracellular matrix (ECM) thickness (μm) of Exophiala dermatitidis mono- (Ed pure) and co-culture biofilms with Pseudomonas aeruginosa (Pa). N = 10.

    Article Snippet: ECM thickness was determined by staining with Invitrogen FilmTracer SYPRO ruby biofilm matrix stain (Thermo Fisher Scientific, Waltham, MA, United States) and subsequent CLSM.

    Techniques: Co-Culture Assay

    Exophiala dermatitidis (Ed) biofilm formation after 48 h at 36°C with culture filtrates of planktonic (plankt) or biofilm (BF) culture from Pseudomonas aeruginosa (Pa). Also, Ed culture filtrate was added. As a control, biofilm formation in pure ASM was assessed. N = 3.

    Journal: Frontiers in Microbiology

    Article Title: Phenotypical Characteristics of the Black Yeast Exophiala dermatitidis Are Affected by Pseudomonas aeruginosa in an Artificial Sputum Medium Mimicking Cystic Fibrosis–Like Conditions

    doi: 10.3389/fmicb.2020.00471

    Figure Lengend Snippet: Exophiala dermatitidis (Ed) biofilm formation after 48 h at 36°C with culture filtrates of planktonic (plankt) or biofilm (BF) culture from Pseudomonas aeruginosa (Pa). Also, Ed culture filtrate was added. As a control, biofilm formation in pure ASM was assessed. N = 3.

    Article Snippet: ECM thickness was determined by staining with Invitrogen FilmTracer SYPRO ruby biofilm matrix stain (Thermo Fisher Scientific, Waltham, MA, United States) and subsequent CLSM.

    Techniques:

    Exophiala dermatitidis (Ed) biofilms formed after 24 h (gray) or 48 h (black) at 36°C in artificial sputum medium (ASM). (A) Biofilms in mono- (Ed pure) and co-culture with Pseudomonas aeruginosa (Pa) wild-type (WT) strains PA07 and PA14, as well as two quorum-sensing (QS) mutants lacking LasR and RhlR. Biofilms were estimated by detachment of biofilm with 0.1% dithiothreitol and subsequent count of colony-forming units (CFU per mL). (B) Biofilms of Exophiala dermatitidis (Ed, P2) and P. aeruginosa (Pa, PA14 WT) after 24 h of incubation at 35°C in ASM treated with N-(3-oxododecanoyl)-L-homoserine lactone (3-oxo-C12-HSL) in a concentration gradient from 0 to 100 μM in DMSO. Biomass in biofilm was measured by staining with 0.1% crystal violet. For correction of biomass in biofilm, difference between the DMSO and the DMSO plus 3-oxo C12 HSL treated biofilms has been estimated and subtracted from the positive control. OD, optical density. Unpaired t -test, * P > 0.05; ** P

    Journal: Frontiers in Microbiology

    Article Title: Phenotypical Characteristics of the Black Yeast Exophiala dermatitidis Are Affected by Pseudomonas aeruginosa in an Artificial Sputum Medium Mimicking Cystic Fibrosis–Like Conditions

    doi: 10.3389/fmicb.2020.00471

    Figure Lengend Snippet: Exophiala dermatitidis (Ed) biofilms formed after 24 h (gray) or 48 h (black) at 36°C in artificial sputum medium (ASM). (A) Biofilms in mono- (Ed pure) and co-culture with Pseudomonas aeruginosa (Pa) wild-type (WT) strains PA07 and PA14, as well as two quorum-sensing (QS) mutants lacking LasR and RhlR. Biofilms were estimated by detachment of biofilm with 0.1% dithiothreitol and subsequent count of colony-forming units (CFU per mL). (B) Biofilms of Exophiala dermatitidis (Ed, P2) and P. aeruginosa (Pa, PA14 WT) after 24 h of incubation at 35°C in ASM treated with N-(3-oxododecanoyl)-L-homoserine lactone (3-oxo-C12-HSL) in a concentration gradient from 0 to 100 μM in DMSO. Biomass in biofilm was measured by staining with 0.1% crystal violet. For correction of biomass in biofilm, difference between the DMSO and the DMSO plus 3-oxo C12 HSL treated biofilms has been estimated and subtracted from the positive control. OD, optical density. Unpaired t -test, * P > 0.05; ** P

    Article Snippet: ECM thickness was determined by staining with Invitrogen FilmTracer SYPRO ruby biofilm matrix stain (Thermo Fisher Scientific, Waltham, MA, United States) and subsequent CLSM.

    Techniques: Co-Culture Assay, Incubation, Concentration Assay, Staining, Positive Control

    (A) Pseudomonas aeruginosa (Pa) and Exophiala dermatitidis (Ed) biofilm formation in monocultures (pure) after 48 h of biofilm formation at 35°C under aerobic (black) or anaerobic (gray) conditions. Biofilms were quantified after biofilm detachment with 0.1% dithiothreitol. (B) Ed biofilm formed after 48 h at 36°C in ASM under hypoxic conditions in monoculture (ED pure) or co-culture with Pa. Biofilms were estimated by detachment of biofilm with 0.1% dithiothreitol and subsequent count of colony-forming units (CFU per mL). Paired t -test, * P

    Journal: Frontiers in Microbiology

    Article Title: Phenotypical Characteristics of the Black Yeast Exophiala dermatitidis Are Affected by Pseudomonas aeruginosa in an Artificial Sputum Medium Mimicking Cystic Fibrosis–Like Conditions

    doi: 10.3389/fmicb.2020.00471

    Figure Lengend Snippet: (A) Pseudomonas aeruginosa (Pa) and Exophiala dermatitidis (Ed) biofilm formation in monocultures (pure) after 48 h of biofilm formation at 35°C under aerobic (black) or anaerobic (gray) conditions. Biofilms were quantified after biofilm detachment with 0.1% dithiothreitol. (B) Ed biofilm formed after 48 h at 36°C in ASM under hypoxic conditions in monoculture (ED pure) or co-culture with Pa. Biofilms were estimated by detachment of biofilm with 0.1% dithiothreitol and subsequent count of colony-forming units (CFU per mL). Paired t -test, * P

    Article Snippet: ECM thickness was determined by staining with Invitrogen FilmTracer SYPRO ruby biofilm matrix stain (Thermo Fisher Scientific, Waltham, MA, United States) and subsequent CLSM.

    Techniques: Co-Culture Assay