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Nat Methods. 2012 Jun 3;9(7):743-8. doi: 10.1038/nmeth.2069.

Single-cell systems biology by super-resolution imaging and combinatorial labeling.

Author information

1
Program in Biochemistry and Molecular Biophysics, California Institute of Technology, Pasadena, California, USA.

Abstract

Fluorescence microscopy is a powerful quantitative tool for exploring regulatory networks in single cells. However, the number of molecular species that can be measured simultaneously is limited by the spectral overlap between fluorophores. Here we demonstrate a simple but general strategy to drastically increase the capacity for multiplex detection of molecules in single cells by using optical super-resolution microscopy (SRM) and combinatorial labeling. As a proof of principle, we labeled mRNAs with unique combinations of fluorophores using fluorescence in situ hybridization (FISH), and resolved the sequences and combinations of fluorophores with SRM. We measured mRNA levels of 32 genes simultaneously in single Saccharomyces cerevisiae cells. These experiments demonstrate that combinatorial labeling and super-resolution imaging of single cells is a natural approach to bring systems biology into single cells.

PMID:
22660740
PMCID:
PMC3418883
DOI:
10.1038/nmeth.2069
[Indexed for MEDLINE]
Free PMC Article

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