Journal: BMC Microbiology

Article Title: Distinct roles of long/short fimbriae and gingipains in homotypic biofilm development by Porphyromonas gingivalis

doi: 10.1186/1471-2180-9-105

Figure Lengend Snippet: Homotypic biofilm formation by P. gingivalis wild-type strain and mutants in PBS . P. gingivalis strains were stained with CFSE (green) and incubated in PBS for 24 hours. After washing, the biofilms that developed on the coverglass were observed with a CLSM equipped with a 40× objective. Optical sections were obtained along the z axis at 0.7-μm intervals, and images of the x - y and x - z planes were reconstructed with an imaging software as described in the text. Upper panels indicate z stacks of the x-y sections. Lower panels are x-z sections. P. gingivalis strains used in this assay are listed in Table 4. The experiment was repeated independently three times with each strain in triplicate. Representative images are shown.

Article Snippet: Quantitative and structural analysis of homotypic P. gingivalis biofilms was accomplished by confocal laser scanning microscopy (CLSM, Radiance 2100, Bio-Rad) and subsequent image analysis [ ].

Techniques: Staining, Incubation, Imaging, Software

Journal: BMC Microbiology

Article Title: Distinct roles of long/short fimbriae and gingipains in homotypic biofilm development by Porphyromonas gingivalis

doi: 10.1186/1471-2180-9-105

Figure Lengend Snippet: Quantification of homotypic biofilms formed by P. gingivalis wild-type strain and mutants in PBS . Biofilms were formed as described in Figure 1, and 10 fields per a sample were randomly recorded and quantified with a CLSM. Z stacks of the x-y sections were converted to composite images to quantify each biovolume as described in the text. Standard error bars are shown. Statistical analysis was performed using a Scheffe test. * p < 0.05 and ** p < 0.01 in comparison to the wild-type strain. P. gingivalis strains used in this assay are listed in Table 4.

Article Snippet: Quantitative and structural analysis of homotypic P. gingivalis biofilms was accomplished by confocal laser scanning microscopy (CLSM, Radiance 2100, Bio-Rad) and subsequent image analysis [ ].

Techniques:

Journal: BMC Microbiology

Article Title: Distinct roles of long/short fimbriae and gingipains in homotypic biofilm development by Porphyromonas gingivalis

doi: 10.1186/1471-2180-9-105

Figure Lengend Snippet: Homotypic biofilm formation by P. gingivalis wild-type strain and mutants in dTSB . P. gingivalis strains were stained with CFSE (green) and incubated in dTSB for 24 hours. After washing, the biofilms that developed on the coverglasses were observed with a CLSM equipped with a 40× objective. Optical sections were obtained along the z axis at 0.7-μm intervals, and images of the x - y and x - z planes were reconstructed with imaging software, as described in the text. Upper panels indicate z stacks of the x-y sections. Lower panels show x-z sections. The experiment was repeated independently three times with each strain in triplicate. Representative images are shown.

Article Snippet: Quantitative and structural analysis of homotypic P. gingivalis biofilms was accomplished by confocal laser scanning microscopy (CLSM, Radiance 2100, Bio-Rad) and subsequent image analysis [ ].

Techniques: Staining, Incubation, Imaging, Software

Journal: BMC Microbiology

Article Title: Distinct roles of long/short fimbriae and gingipains in homotypic biofilm development by Porphyromonas gingivalis

doi: 10.1186/1471-2180-9-105

Figure Lengend Snippet: Exopolysaccharide production by P. gingivalis wild-type strain and mutants in dTSB . A) Visualization of exopolysaccharide production in biofilms formed by P. gingivalis strains after staining with FITC-labelled concanavalin A and wheat germ agglutinin (green). Bacteria were stained with DAPI (blue). Fluorescent images were obtained using a CLSM. The z stack of the x-y sections was converted to composite images with the "Volume" function using Imaris software, after which a y stack of the x-z sections was created and is presented here. B) Fluorescent images were quantified using Imaris software and average of total exopolysaccharide biovolume per field was calculated. C) Exopolysaccharide levels are expressed as the ratio of exopolysaccharide/cells (FITC/DAPI) fluorescence. The experiment was repeated independently three times. Data are presented as averages of 8 fields per sample with standard errors of the means. Statistical analysis was performed using a Scheffe test. * p < 0.05 and ** p < 0.01 in comparison to the wild-type strain.

Article Snippet: Quantitative and structural analysis of homotypic P. gingivalis biofilms was accomplished by confocal laser scanning microscopy (CLSM, Radiance 2100, Bio-Rad) and subsequent image analysis [ ].

Techniques: Staining, Software, Fluorescence