Protocol to perform dynamic microfluidic single-cell cultivation of C. glutamicum

Luisa Blöbaum; Sarah Täuber; Alexander Grünberger

doi:10.1016/j.xpro.2023.102436

Protocol to perform dynamic microfluidic single-cell cultivation of C. glutamicum

STAR Protoc. 2023 Sep 15;4(3):102436. doi: 10.1016/j.xpro.2023.102436. Epub 2023 Aug 6.

Authors

Luisa Blöbaum¹, Sarah Täuber², Alexander Grünberger³

Affiliations

¹ Multiscale Bioengineering, Technical Faculty, Bielefeld University, 33615 Bielefeld, Germany; Center for Biotechnology (CeBiTec), Bielefeld University, 33615 Bielefeld, Germany. Electronic address: luisa.bloebaum@uni-bielefeld.de.
² Multiscale Bioengineering, Technical Faculty, Bielefeld University, 33615 Bielefeld, Germany; Center for Biotechnology (CeBiTec), Bielefeld University, 33615 Bielefeld, Germany.
³ Multiscale Bioengineering, Technical Faculty, Bielefeld University, 33615 Bielefeld, Germany; Microsystems in Bioprocess Engineering, Institute of Process Engineering in Life Sciences, Karlsruhe Institute of Technology, 76131 Karlsruhe, Germany. Electronic address: alexander.gruenberger@kit.edu.

Abstract

Here, we present a protocol for the design, fabrication, and usage of a polydimethylsiloxane (PDMS)-based chip for dynamic microfluidic single-cell cultivation of Corynebacterium glutamicum. We describe steps for flow profile establishment and biological preparation. We then detail time-lapse imaging to observe reactions of C. glutamicum to repeated environmental changes in the range of seconds. This system can be adapted to other organisms with a cell wall and soluble non-gaseous environmental factors like nutrients. For complete details on the use and execution of this protocol, please refer to Täuber et al..¹.

Keywords: Biotechnology and bioengineering; Microbiology; Microscopy; Single Cell.

MeSH terms

Corynebacterium glutamicum*
Microfluidics*