Monitoring of dynamic microbiological processes using real-time flow cytometry.
We describe a straightforward approach to continuously monitor a variety of highly dynamic microbiological processes in millisecond resolution with flow cytometry, using standard bench-top instrumentation. Four main experimental examples are provided, namely: (1) green fluorescent protein expression by antibiotic-stressed Escherichia coli, (2) fluorescent labeling of heat-induced membrane damage in an autochthonous freshwater bacterial community, (3) the initial growth response of late stationary E. coli cells inoculated into fresh growth media, and (4) oxidative disinfection of a mixed culture of auto-fluorescent microorganisms. These examples demonstrate the broad applicability of the method to diverse biological experiments, showing that it allows the collection of detailed, time-resolved information on complex processes.
Department of Environmental Microbiology, Eawag - Swiss Federal Institute for Aquatic Science and Technology, Dübendorf, Switzerland ; Department of Environmental Systems Science, ETH Zürich, Zürich, Switzerland.
Markus
M
Arnoldini
Tobias
T
Heck
Alfonso
A
Blanco-Fernández
Frederik
F
Hammes
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D052160:Benzothiazoles; D002462:Cell Membrane / Q000187:drug effects; D016825:Chlamydomonas reinhardtii / Q000235:genetics / Q000378:metabolism*; D002939:Ciprofloxacin / Q000494:pharmacology; D003959:Diamines; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D005434:Flow Cytometry / Q000379:methods*; D005452:Fluoresceins / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D015964:Gene Expression Regulation, Bacterial; D018506:Gene Expression Regulation, Plant; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006358:Hot Temperature; D046931:Microcystis / Q000235:genetics / Q000378:metabolism*; D009930:Organic Chemicals / Q000737:chemistry; D011804:Quinolines; D011938:Rec A Recombinases / Q000235:genetics / Q000378:metabolism; D013312:Stress, Physiological; D013997:Time Factors; D014158:Transcription, Genetic / Q000187:drug effects
D000900:Anti-Bacterial Agents; D052160:Benzothiazoles; D003959:Diamines; D005452:Fluoresceins; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; C027780:carboxyfluoresceindiacetate; D049452:Green Fluorescent Proteins; C098022:SYBR Green I; D002939:Ciprofloxacin; D011938:Rec A Recombinases