A microscope automated fluidic system to study bacterial processes in real time.
Most time lapse microscopy experiments studying bacterial processes ie growth, progression through the cell cycle and motility have been performed on thin nutrient agar pads. An important limitation of this approach is that dynamic perturbations of the experimental conditions cannot be easily performed. In eukaryotic cell biology, fluidic approaches have been largely used to study the impact of rapid environmental perturbations on live cells and in real time. However, all these approaches are not easily applicable to bacterial cells because the substrata are in all cases specific and also because microfluidics nanotechnology requires a complex lithography for the study of micrometer sized bacterial cells. In fact, in many cases agar is the experimental solid substratum on which bacteria can move or even grow. For these reasons, we designed a novel hybrid micro fluidic device that combines a thin agar pad and a custom flow chamber. By studying several examples, we show that this system allows real time analysis of a broad array of biological processes such as growth, development and motility. Thus, the flow chamber system will be an essential tool to study any process that take place on an agar surface at the single cell level.
Ducret
Adrien
A
Aix Marseille Université, Laboratoire de Chimie Bactérienne (UPR 9043), Institut de Microbiologie de la Méditerranée (IFR 88), CNRS, 31, Chemin Joseph Aiguier, Marseille, France.
Maisonneuve
Etienne
E
Notareschi
Philippe
P
Grossi
Alain
A
Mignot
Tâm
T
Dukan
Sam
S
D000362:Agar / Q000737:chemistry; D001331:Automation; D001419:Bacteria / Q000378:metabolism*; D018407:Bacterial Physiological Phenomena; D018929:Cell Culture Techniques / Q000295:instrumentation; D002453:Cell Cycle; D004867:Equipment Design; D004926:Escherichia coli / Q000378:metabolism; D046210:Microfluidic Analytical Techniques; D044085:Microfluidics / Q000295:instrumentation; D008853:Microscopy / Q000295:instrumentation* / Q000379:methods; D016940:Myxococcus / Q000378:metabolism; D036103:Nanotechnology / Q000379:methods
D000362:Agar