Microfluidic single-cell real-time PCR for comparative analysis of gene expression patterns

Veronica Sanchez-Freire; Antje D. Ebert; Tomer Kalisky; Stephen R. Quake; Joseph C. Wu

doi:10.1038/nprot.2012.021

Microfluidic single-cell real-time PCR for comparative analysis of gene expression patterns

Veronica Sanchez-Freire, Antje D. Ebert, Tomer Kalisky, Stephen R. Quake, Joseph C. Wu

Stanford University

Research output: Contribution to journal › Article › peer-review

139 Scopus citations

Abstract

Single-cell quantitative real-time PCR (qRT-PCR) combined with high-throughput arrays allows the analysis of gene expression profiles at a molecular level in approximately 11 h after cell sample collection. We present here a high-content microfluidic real-time platform as a powerful tool for comparatively investigating the regulation of developmental processes in single cells. This approach overcomes the limitations involving heterogeneous cell populations and sample amounts, and may shed light on differential regulation of gene expression in normal versus disease-related contexts. Furthermore, high-throughput single-cell qRT-PCR provides a standardized, comparative assay for in-depth analysis of the mechanisms underlying human pluripotent stem cell self-renewal and differentiation.

Original language	English
Pages (from-to)	829-838
Number of pages	10
Journal	Nature Protocols
Volume	7
Issue number	5
DOIs	https://doi.org/10.1038/nprot.2012.021
State	Published - 5 Apr 2012
Externally published	Yes

Bibliographical note

Funding Information:
acknoWleDGMents We are grateful to P.E. de Almeida for FACS discussion. We acknowledge funding support from the Swiss National Science Foundation PBBEP3_129803 (V.S.-F.); the German Research Foundation (A.D.E.); the Howard Hughes Medical Institute (S.R.Q.); the US National Institutes of Health (NIH) DP2OD004437, RC1AG036142, R01AI085575 (J.C.W.), and the Burroughs Wellcome Foundation (J.C.W.).

Funding

acknoWleDGMents We are grateful to P.E. de Almeida for FACS discussion. We acknowledge funding support from the Swiss National Science Foundation PBBEP3_129803 (V.S.-F.); the German Research Foundation (A.D.E.); the Howard Hughes Medical Institute (S.R.Q.); the US National Institutes of Health (NIH) DP2OD004437, RC1AG036142, R01AI085575 (J.C.W.), and the Burroughs Wellcome Foundation (J.C.W.).

Funders	Funder number
US National Institutes of Health
National Institutes of Health	DP2OD004437, R01AI085575
Howard Hughes Medical Institute
National Institute on Aging	RC1AG036142
Burroughs Wellcome Fund
Deutsche Forschungsgemeinschaft
Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung	PBBEP3_129803

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1038/nprot.2012.021

Cite this

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Microfluidic single-cell real-time PCR for comparative analysis of gene expression patterns

Abstract

Bibliographical note

Funding

UN SDGs

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