algorithms nis elements viewer software nikon version 5 21 00 r software  (Nikon)

Journal: PLOS Pathogens

Article Title: Pseudomonas aeruginosa senses and responds to epithelial potassium flux via Kdp operon to promote biofilm

doi: 10.1371/journal.ppat.1011453

Figure Lengend Snippet: (A and B) CFBE41o - cells on glass coverslips are infected with a 1:1:1 mixture of TFP-, YFP-, and tdTomato-producing P . aeruginosa while treated with NS19504 (25 μM) or 0.05% DMSO in respiratory epithelial co-culture biofilm assay and imaged by fluorescent microscopy at 1-, 3-, and 6-hours post-infection. (A) Representative images of bacterial coalescence at 1-, 3-, and 6-hours growth. Scale bar represents 10 μm. White box indicates area of magnification in panel B. (B) Magnification of one quarter of panel A images demonstrating multicolor aggregates (polyclonal) compared to single color aggregates (monoclonal). (C) Proportion of colocalized bacteria at 6 hours measured with Nikon Elements Software. Bar represents mean with dots representing each of the four biologic replicates and error bars represent standard error of the mean. Statistical significance was determined by unpaired t-test (* p<0.05).

Article Snippet: Bacterial biomass is calculated using the Nikon Elements software algorithm that quantifies volumes of thresholded objects in the images.

Techniques: Infection, Co-Culture Assay, Biofilm Production Assay, Microscopy, Bacteria, Software

Journal: PLOS Pathogens

Article Title: Pseudomonas aeruginosa senses and responds to epithelial potassium flux via Kdp operon to promote biofilm

doi: 10.1371/journal.ppat.1011453

Figure Lengend Snippet: (A) CFBE41o - cells on glass coverslips were infected with GFP-producing P . aeruginosa (green) while treated with paxilline (10 μM), a BK Ca channel inhibitor, or DMSO at 0.05% in MEM without phenol red and imaged by fluorescent microscopy at 1, 3, and 6 hours. CFBE41o - cell nuclei are stained with Hoescht33342 (blue). Scale bar represents 10 μm. (B) Biomass (μm 3 /μm 2 ) measurements (measured with Nikon Elements Software) at 1-, 3-, and 6-hours post-inoculation from three independent experiments. Line in bar represents mean value and error bars represent standard error of the mean. Statistical significance was tested by two-way ANOVA with multiple comparisons (*** p<0.001). (C) P . aeruginosa biofilms grown in 96-well plates in LB and SCFM with or without paxilline measured using crystal violet absorbance at 550 nm. (D) Planktonic growth kinetics of P . aeruginosa grown in LB Lennox broth (LB), minimal essential media (MEM), and synthetic cystic fibrosis sputum media (SCFM) with or without paxilline. (E) Biomass (μm 3 /μm 2 ) measurements before and after paxilline treatment. After biofilms were formed and measured on DMSO treated cells for 6 hours, paxilline containing media was introduced for 30 minutes and biofilms were measured. Line represents mean and error bars represent standard error of the mean. Line connecting data points indicates data points from same biologic replicate.

Article Snippet: Bacterial biomass is calculated using the Nikon Elements software algorithm that quantifies volumes of thresholded objects in the images.

Techniques: Infection, Microscopy, Staining, Software