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Cultivation and quantitative single‐cell analysis of Saccharomyces cerevisiae on a multifunctional microfluidic device
Author(s) -
Stratz Simone,
Verboket Pascal Emilio,
Hasler Karina,
Dittrich Petra Stephanie
Publication year - 2018
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201700253
Subject(s) - saccharomyces cerevisiae , yeast , microfluidics , biomolecule , single cell analysis , cell , biophysics , chemistry , intracellular , nanotechnology , materials science , biochemistry , biology
Here, we present a multifunctional microfluidic device whose integrative design enables to combine cell culture studies and quantitative single cell biomolecule analysis. The platform consists of 32 analysis units providing two key features; first, a micrometer‐sized trap for hydrodynamic capture of a single Saccharomyces cerevisiae ( S. cerevisiae ) yeast cell; second, a convenient double‐valve configuration surrounding the trap. Actuating of the outer valve with integrated opening results in a partial isolation in a volume of 11.8 nL, i.e. the cell surrounding fluid can be exchanged diffusion‐based without causing shear stress or cell loss. Actuation of the inner ring‐shaped valve isolates the trapped cell completely in a small analysis volume of 230 pL. The device was used to determine the growth rate of yeast cells ( S. cerevisiae ) under under optimum and oxidative stress conditions. In addition, we successfully quantified the cofactor beta‐nicotinamide adenine dinucleotide phosphate (NAD(P)H) in single and few cells exposed to the different microenvironments. In conclusion, the microdevice enables to analyze the influence of an external stress factor on the cellular fitness in a fast and more comprehensive way as cell growth and intracellular biomolecule levels can be investigated.