UCSD JSOE JSOE

Turning Single Cells Intro Microarrays by Super-Resolution Barcoding

Fri, 11/30/2012 - 2:00pm
Fung Auditorium, Powell-Focht Bioengineering Hall

Long Cai, PhD
Assistant Professor, Chemistry
California Institute of Technology

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.

Biosketch

Long studied chemistry and physics as an undergraduate at Harvard, where he worked for Dudley Herschbach and Bretislav Friedrich on "pendular state" molecules generated by short laser pulses. Long continued graduate studies at Harvard with Sunney Xie in single molecule spectroscopy, where he worked on watching gene expression and decision-making events in single cells with single molecule resolution. He then came to Caltech and became a Beckman Institute fellow in the laboratory of Michael Elowitz in 2006, where he worked on signaling in single yeast cells. Long started his laboratory in the division of Chemistry and Chemical Engineering at Caltech on July 1st, 2010.

 

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