Spatiotemporal microbial single-cell analysis using a high-throughput microfluidics cultivation platform.
Cell-to-cell heterogeneity typically evolves due to a manifold of biological and environmental factors and special phenotypes are often relevant for the fate of the whole population but challenging to detect during conventional analysis. We demonstrate a microfluidic single-cell cultivation platform that incorporates several hundred growth chambers, in which isogenic bacteria microcolonies growing in cell monolayers are tracked by automated time-lapse microscopy with spatiotemporal resolution. The device was not explicitly developed for a specific organism, but has a very generic configuration suitable for various different microbial organisms. In the present study, we analyzed Corynebacterium glutamicum microcolonies, thereby generating complete lineage trees and detailed single-cell data on division behavior and morphology in order to demonstrate the platform's overall capabilities. Furthermore, the occurrence of spontaneously induced stress in individual C. glutamicum cells was investigated by analyzing strains with genetically encoded reporter systems and optically visualizing SOS response. The experiments revealed spontaneous SOS induction in the absence of any external trigger comparable to results obtained by flow cytometry (FC) analyzing cell samples from conventional shake flask cultivation. Our microfluidic setup delivers detailed single-cell data with spatial and temporal resolution; complementary information to conventional FC results.
Forschungszentrum Jülich GmbH, IBG-1: Biotechnology, Jülich, 52425, Germany.
Alexander
A
Grünberger
Forschungszentrum Jülich GmbH, IBG-1: Biotechnology, Jülich, 52425, Germany.
Christopher
C
Probst
Forschungszentrum Jülich GmbH, IBG-1: Biotechnology, Jülich, 52425, Germany.
Stefan
S
Helfrich
Forschungszentrum Jülich GmbH, IBG-1: Biotechnology, Jülich, 52425, Germany.
Arun
A
Nanda
Forschungszentrum Jülich GmbH, IBG-1: Biotechnology, Jülich, 52425, Germany.
Birgit
B
Stute
Forschungszentrum Jülich GmbH, IBG-1: Biotechnology, Jülich, 52425, Germany.
Wolfgang
W
Wiechert
Forschungszentrum Jülich GmbH, IBG-1: Biotechnology, Jülich, 52425, Germany.
Eric
E
von Lieres
Forschungszentrum Jülich GmbH, IBG-1: Biotechnology, Jülich, 52425, Germany.
Katharina
K
Nöh
Forschungszentrum Jülich GmbH, IBG-1: Biotechnology, Jülich, 52425, Germany.
Julia
J
Frunzke
Forschungszentrum Jülich GmbH, IBG-1: Biotechnology, Jülich, 52425, Germany.
Dietrich
D
Kohlheyer
D048230:Corynebacterium glutamicum / Q000166:cytology* / Q000254:growth & development; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods*; D057446:Hydrodynamics; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D012212:Rheology; D013014:SOS Response, Genetics; D059010:Single-Cell Analysis / Q000295:instrumentation / Q000379:methods*; D062211:Spatio-Temporal Analysis
D004129:Dimethylpolysiloxanes
C. glutamicum; microfluidics; single-cell analysis; time-lapse microscopy