Journal: Scientific Reports

Article Title: ciaR impacts biofilm formation by regulating an arginine biosynthesis pathway in Streptococcus sanguinis SK36

doi: 10.1038/s41598-017-17383-1

Figure Lengend Snippet: The impact of ciaR mutation on biofilm formation. ( A ) The biofilms of WT, Δ ciaR and Δ ciaR / ciaR were cultured in a 96-well plate in BM media for 24 hours. Biomass was measured by crystal violet staining. ( B ) Biofilms of WT and Δ ciaR cultured in 4-well chambers after being washed with PBS buffer. ( C ) Biofilms grown in 4-well chambers were stained by SYTO9 and propidium iodide. Fluorescence (left) and differential interference microscopy images (middle) were obtained by confocal laser scanning microscopy. Fluorescence images were analyzed by COMSTAT script, and heat maps of biofilm thickness were generated, which showed the distribution of biomass in biofilms (right). ( D ) The fluorescence images were analyzed by COMSTAT. Biofilm biomass, average thickness, propidium iodide signal and roughness coefficient were quantified, respectively. All the data in Fig. 1D were compared with their WT control. *P ≤ 0.05, **P ≤ 0.01, Student’s t- test. Means and standard deviations from triplicate experiments are shown.

Article Snippet: These biofilms were subsequently observed by confocal laser scanning microscopy (CLSM) and quantified using a COMSTAT script in Matlab software .

Techniques: Mutagenesis, Cell Culture, Staining, Fluorescence, Microscopy, Confocal Laser Scanning Microscopy, Generated, Control

Journal: Scientific Reports

Article Title: ciaR impacts biofilm formation by regulating an arginine biosynthesis pathway in Streptococcus sanguinis SK36

doi: 10.1038/s41598-017-17383-1

Figure Lengend Snippet: Biofilm formation of WT and Δ ciaR under flow cell conditions. Strains were marked by different fluorescent reporters and then cultured in a flow cell system. Image in Fig. 5C was obtained by CLSM. Others were recorded by the fluorescence microscopy. Biofilm biomass was quantified by COMSTAT software. ( A ) The biofilms biomass of WT and Δ ciaR in the flow cell system at different time points. ( B ) WT and Δ ciaR were co-cultured in the flow cell channel and biofilm biomass was measured. ( C ) After being co-cultured for 4 days, the structure of biofilm formed by WT – Δ ciaR was examined by CLSM. For each sample in Fig. 5A and B, ten images from fluorescence microscopy were obtained to calculate the means and standard deviations. *P ≤ 0.05, **P ≤ 0.01, Student’s t- test.

Article Snippet: These biofilms were subsequently observed by confocal laser scanning microscopy (CLSM) and quantified using a COMSTAT script in Matlab software .

Techniques: Cell Culture, Fluorescence, Microscopy, Software