Single-cell Systems Biology by Super-resolution Imaging and Combinatorial Labeling

Overview

Journal Nat Methods

Specialties Biomedical Engineering
Pathology

Date 2012 Jun 5

PMID 22660740

Citations 217

Authors

Eric Lubeck

Long Cai

Affiliations

Soon will be listed here.

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.

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