Cultivation and quantitative single-cell analysis of Saccharomyces cerevisiae on a multifunctional microfluidic device. 2018

Simone Stratz, and Pascal Emilio Verboket, and Karina Hasler, and Petra Stephanie Dittrich
ETH Zurich, Department of Chemistry and Applied Biosciences, Zurich, Switzerland.

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.

UI MeSH Term Description Entries
D009249 NADP Nicotinamide adenine dinucleotide phosphate. A coenzyme composed of ribosylnicotinamide 5'-phosphate (NMN) coupled by pyrophosphate linkage to the 5'-phosphate adenosine 2',5'-bisphosphate. It serves as an electron carrier in a number of reactions, being alternately oxidized (NADP+) and reduced (NADPH). (Dorland, 27th ed) Coenzyme II,Nicotinamide-Adenine Dinucleotide Phosphate,Triphosphopyridine Nucleotide,NADPH,Dinucleotide Phosphate, Nicotinamide-Adenine,Nicotinamide Adenine Dinucleotide Phosphate,Nucleotide, Triphosphopyridine,Phosphate, Nicotinamide-Adenine Dinucleotide
D010084 Oxidation-Reduction A chemical reaction in which an electron is transferred from one molecule to another. The electron-donating molecule is the reducing agent or reductant; the electron-accepting molecule is the oxidizing agent or oxidant. Reducing and oxidizing agents function as conjugate reductant-oxidant pairs or redox pairs (Lehninger, Principles of Biochemistry, 1982, p471). Redox,Oxidation Reduction
D004129 Dimethylpolysiloxanes Silicone polymers which consist of silicon atoms substituted with methyl groups and linked by oxygen atoms. They comprise a series of biocompatible materials used as liquids, gels or solids; as film for artificial membranes, gels for implants, and liquids for drug vehicles; and as antifoaming agents. Dimethylsiloxanes,Polydimethylsiloxanes,Dimethylpolysiloxane,Dimethylsiloxane
D012441 Saccharomyces cerevisiae A species of the genus SACCHAROMYCES, family Saccharomycetaceae, order Saccharomycetales, known as "baker's" or "brewer's" yeast. The dried form is used as a dietary supplement. Baker's Yeast,Brewer's Yeast,Candida robusta,S. cerevisiae,Saccharomyces capensis,Saccharomyces italicus,Saccharomyces oviformis,Saccharomyces uvarum var. melibiosus,Yeast, Baker's,Yeast, Brewer's,Baker Yeast,S cerevisiae,Baker's Yeasts,Yeast, Baker
D046210 Microfluidic Analytical Techniques Methods utilizing the principles of MICROFLUIDICS for sample handling, reagent mixing, and separation and detection of specific components in fluids. Microfluidic Analysis,Analyses, Microfluidic,Analysis, Microfluidic,Analytical Technique, Microfluidic,Analytical Techniques, Microfluidic,Microfluidic Analyses,Microfluidic Analytical Technique,Technique, Microfluidic Analytical,Techniques, Microfluidic Analytical
D056656 Lab-On-A-Chip Devices Microdevices that combine microfluidics technology with electrical and/or mechanical functions for analyzing very small fluid volumes. They consist of microchannels etched into substrates made of silicon, glass, or polymer using processes similar to photolithography. The test fluids in the channels can then interact with different elements such as electrodes, photodetectors, chemical sensors, pumps, and valves. Microchip Analytical Devices,Microfluidic Devices,Microfluidic Lab-On-A-Chip,Microfluidic Microchips,Nanochip Analytical Devices,Analytical Device, Microchip,Analytical Device, Nanochip,Analytical Devices, Microchip,Analytical Devices, Nanochip,Device, Lab-On-A-Chip,Device, Microchip Analytical,Device, Microfluidic,Device, Nanochip Analytical,Devices, Lab-On-A-Chip,Devices, Microchip Analytical,Devices, Microfluidic,Devices, Nanochip Analytical,Lab On A Chip Devices,Lab-On-A-Chip Device,Lab-On-A-Chip, Microfluidic,Lab-On-A-Chips, Microfluidic,Microchip Analytical Device,Microchip, Microfluidic,Microchips, Microfluidic,Microfluidic Device,Microfluidic Lab On A Chip,Microfluidic Lab-On-A-Chips,Microfluidic Microchip,Nanochip Analytical Device
D057446 Hydrodynamics The motion of fluids, especially noncompressible liquids, under the influence of internal and external forces. Fluid Dynamics,Dynamic, Fluid,Dynamics, Fluid,Fluid Dynamic,Hydrodynamic
D058948 Cell Tracking Non-invasive imaging of cells that have been labeled non-destructively, such as with nanoemulsions or reporter genes that can be detected by molecular imaging, to monitor their location, viability, cell lineage expansion, response to drugs, movement, or other behaviors in vivo. Cell Trackings,Tracking, Cell,Trackings, Cell
D059010 Single-Cell Analysis Assaying the products of or monitoring various biochemical processes and reactions in an individual cell. Analyses, Single-Cell,Analysis, Single-Cell,Single Cell Analysis,Single-Cell Analyses
D018929 Cell Culture Techniques Methods for maintaining or growing CELLS in vitro. Cell Culture,Cell Culture Technique,Cell Cultures,Culture Technique, Cell,Culture Techniques, Cell

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