title: Imaging and analysis of Pseudomonas aeruginosa swarming and rhamnolipid production.
abstract: Many bacteria spread over surfaces by "swarming" in groups. A problem for scientists who study swarming is the acquisition of statistically significant data that distinguish two observations or detail the temporal patterns and two-dimensional heterogeneities that occur. It is currently difficult to quantify differences between observed swarm phenotypes. Here, we present a method for acquisition of temporal surface motility data using time-lapse fluorescence and bioluminescence imaging. We specifically demonstrate three applications of our technique with the bacterium Pseudomonas aeruginosa. First, we quantify the temporal distribution of P. aeruginosa cells tagged with green fluorescent protein (GFP) and the surfactant rhamnolipid stained with the lipid dye Nile red. Second, we distinguish swarming of P. aeruginosa and Salmonella enterica serovar Typhimurium in a coswarming experiment. Lastly, we quantify differences in swarming and rhamnolipid production of several P. aeruginosa strains. While the best swarming strains produced the most rhamnolipid on surfaces, planktonic culture rhamnolipid production did not correlate with surface growth rhamnolipid production.
authors: Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana 46556, USA.
authors: Joshua D
authors: JD
authors: Morris
authors: Jessica L
authors: JL
authors: Hewitt
authors: Lawrence G
authors: LG
authors: Wolfe
authors: Nachiket G
authors: NG
authors: Kamatkar
authors: Sarah M
authors: SM
authors: Chapman
authors: Justin M
authors: JM
authors: Diener
authors: Andrew J
authors: AJ
authors: Courtney
authors: W Matthew
authors: WM
authors: Leevy
authors: Joshua D
authors: JD
authors: Shrout
mesh_terms: D017930:Genes, Reporter; D006017:Glycolipids / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008124:Locomotion; D049449:Luminescence; D011550:Pseudomonas aeruginosa / Q000378:metabolism / Q000502:physiology*; D012486:Salmonella typhimurium / Q000378:metabolism / Q000502:physiology; D013194:Staining and Labeling / Q000379:methods; D059008:Time-Lapse Imaging / Q000379:methods*
chemical_list: D006017:Glycolipids; C418382:rhamnolipid; D049452:Green Fluorescent Proteins