Issue 14, 2015

Capture and enumeration of mRNA transcripts from single cells using a microfluidic device

Abstract

Accurate measurement of RNA transcripts from single cells will enable the precise classification of cell types and characterization of the heterogeneity in cell populations that play key roles in normal cellular physiology and diseases. As a step towards this end, we have developed a microfluidic device and methods for automatic hydrodynamic capture of single mammalian cells and subsequent immobilization and digital counting of polyadenylated mRNA molecules released from the individual cells. Using single-molecule fluorescence imaging, we have demonstrated that polyadenylated mRNA molecules from single HeLa cells can be captured within minutes by hybridization to polydeoxyribothymidine oligonucleotides covalently attached on the glass surface in the device. The total mRNA molecule counts in the individual HeLa cells are found to vary significantly from one another. Our technology opens up the possibility of direct digital enumeration of RNA transcripts from single cells with single-molecule sensitivity using a single integrated microfluidic device.

Graphical abstract: Capture and enumeration of mRNA transcripts from single cells using a microfluidic device

Supplementary files

Article information

Article type
Paper
Submitted
19 Apr 2015
Accepted
27 May 2015
First published
27 May 2015

Lab Chip, 2015,15, 2968-2980

Author version available

Capture and enumeration of mRNA transcripts from single cells using a microfluidic device

M. T. Walsh, A. P. Hsiao, H. S. Lee, Z. Liu and X. Huang, Lab Chip, 2015, 15, 2968 DOI: 10.1039/C5LC00445D

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