Flow-based microfluidic device for quantifying bacterial chemotaxis in stable, competing gradients.
Chemotaxis is the migration of cells in gradients of chemoeffector molecules. Although multiple, competing gradients must often coexist in nature, conventional approaches for investigating bacterial chemotaxis are suboptimal for quantifying migration in response to gradients of multiple signals. In this work, we developed a microfluidic device for generating precise and stable gradients of signaling molecules. We used the device to investigate the effects of individual and combined chemoeffector gradients on Escherichia coli chemotaxis. Laminar flow-based diffusive mixing was used to generate gradients, and the chemotactic responses of cells expressing green fluorescent protein were determined using fluorescence microscopy. Quantification of the migration profiles indicated that E. coli was attracted to the quorum-sensing molecule autoinducer-2 (AI-2) but was repelled from the stationary-phase signal indole. Cells also migrated toward higher concentrations of isatin (indole-2,3-dione), an oxidized derivative of indole. Attraction to AI-2 overcame repulsion by indole in equal, competing gradients. Our data suggest that concentration-dependent interactions between attractant and repellent signals may be important determinants of bacterial colonization of the gut.
Englert
Derek L
DL
Artie McFerrin Department of Chemical Engineering, Texas A&M University, College Station, 77843-3122, USA.
Manson
Michael D
MD
Jayaraman
Arul
A
D001431:Bacteriological Techniques / Q000379:methods*; D002633:Chemotaxis; D004926:Escherichia coli / Q000502:physiology*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006714:Homoserine / Q000031:analogs & derivatives / Q000378:metabolism; D007211:Indoles / Q000378:metabolism; D007510:Isatin / Q000378:metabolism; D007783:Lactones / Q000378:metabolism; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D008856:Microscopy, Fluorescence
D007211:Indoles; D007783:Lactones; C091088:N-octanoylhomoserine lactone; D049452:Green Fluorescent Proteins; D006714:Homoserine; D007510:Isatin; C030374:indole