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Year: 2013 Issue: 82

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Year: 2013 Issue: 80

A microfluidic chip for the versatile chemical analysis of single cells.

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A microfluidic chip for the versatile chemical analysis of single cells.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Eyer, Klaus Kuhn, Phillip Stratz, Simone Dittrich, Petra S.
Description	Country affiliation: Switzerland Author Affiliation: Eyer K ( Department of Chemistry and Applied Biosciences, ETH Zurich, Switzerland.)
Abstract	We present a microfluidic device that enables the quantitative determination of intracellular biomolecules in multiple single cells in parallel. For this purpose, the cells are passively trapped in the middle of a microchamber. Upon activation of the control layer, the cell is isolated from the surrounding volume in a small chamber. The surrounding volume can then be exchanged without affecting the isolated cell. However, upon short opening and closing of the chamber, the solution in the chamber can be replaced within a few hundred milliseconds. Due to the reversibility of the chambers, the cells can be exposed to different solutions sequentially in a highly controllable fashion, e.g. for incubation, washing, and finally, cell lysis. The tightly sealed microchambers enable the retention of the lysate, minimize and control the dilution after cell lysis. Since lysis and analysis occur at the same location, high sensitivity is retained because no further dilution or loss of the analytes occurs during transport. The microchamber design therefore enables the reliable and reproducible analysis of very small copy numbers of intracellular molecules (attomoles, zeptomoles) released from individual cells. Furthermore, many microchambers can be arranged in an array format, allowing the analysis of many cells at once, given that suitable optical instruments are used for monitoring. We have already used the platform for proof-of-concept studies to analyze intracellular proteins, enzymes, cofactors and second messengers in either relative or absolute quantifiable manner.
File Format	HTM / HTML
e-ISSN	1940087X
DOI	10.3791/50618
Journal	Journal of Visualized Experiments
Issue Number	80
Language	English
Publisher	MyJove Corp.
Publisher Date	2013-10-15
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Physical Sciences Discipline Life Sciences Discipline Medicine Microfluidic Analytical Techniques Instrumentation Single-cell Analysis Dimethylpolysiloxanes Chemistry Equipment Design Hek293 Cells Silicon U937 Cells Research Support, Non-u.s. Gov't Video-audio Media
Content Type	Text
Resource Type	Article

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