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Yeast. 2006 Oct-Nov;23(14-15):1065-73.

Microfluidics device for single cell gene expression analysis in Saccharomyces cerevisiae.

Author information

  • 1University of Texas Health Science Center at San Antonio, Sam and Ann Barshop Institute for Longevity and Ageing Studies, Department of Cellular and Structural Biology, San Antonio, TX 78245-3207, USA. james@ryley.com

Abstract

We have measured single-cell gene expression over time using a microfluidics-based flow cell which physically traps individual yeast using microm-sized structures (yeast jails). Our goal was to determine variability of gene expression within a cell over time, as well as variability between individual cells. In our flow cell system, yeast jails are fabricated out of PDMS and gene expression is visualized using fluorescently-tagged proteins of interest. Previously, single-cell yeast work has been done using micromanipulation on agar, or FACS. In the present device agar is eliminated, resulting in a superior optical system. The flow of media through the flow cell washes daughter cells away, eliminating the need for micromanipulation. Unlike FACS, the described device can track individual yeast over a time course of many hours. The flow cells are compatible with the needs of quantitative fluorescence microscopy, and allow simultaneous measurements to be done on a large number of individual yeast. We used these flow cells to determine the expression of HSP104-GFPand RAS2-YFP, genes known to affect yeast life span. The results demonstrate inter-cell variation in expression of both genes that could not have been detected without this single-cell analysis.

Copyright (c) 2006 John Wiley & Sons, Ltd.

PMID:
17083143
[PubMed - indexed for MEDLINE]
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