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Imaging Chromosome Structure in Bacteria by Super-Resolution Microscopy

Cai, Long (2014) Imaging Chromosome Structure in Bacteria by Super-Resolution Microscopy. Biophysical Journal, 106 (2). 375A. ISSN 0006-3495. doi:10.1016/j.bpj.2013.11.2120.

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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 separability of 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. The basis for this new approach are the following: given the 10 nanometers resolution of a super-resolution microscope and a typical cell a size of (10um)3, individual cells contains effectively 109 super-resolution pixels or bits of information. Most eukaryotic cells have 104 genes and cellular abundances of 10-100 copies per transcript. Thus, under a super-resolution microscope, an individual cell has 1000 times more pixel volume or information capacities than is needed to encode all transcripts within that cell. 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 the mRNA levels of 32 genes simultaneously in single cells. In addition, we have performed DNA-FISH experiments simultaneously with RNA-FISH to image both the chromosome structure and transcription in single E.coli cells.

Item Type:Article
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Cai, Long0000-0002-7154-5361
Additional Information:© 2014 Biophysical Society. Published by Elsevier Inc.
Issue or Number:2
Record Number:CaltechAUTHORS:20140710-095942704
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Official Citation:Long Cai, Imaging Chromosome Structure in Bacteria by Super-Resolution Microscopy, Biophysical Journal, Volume 106, Issue 2, Supplement 1, 28 January 2014, Page 375a, ISSN 0006-3495, (
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:47134
Deposited By: Tony Diaz
Deposited On:10 Jul 2014 19:34
Last Modified:10 Nov 2021 17:35

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