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Published January 28, 2014 | public
Journal Article

Imaging Chromosome Structure in Bacteria by Super-Resolution Microscopy

Cai, Long ORCID icon

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 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.

Additional Information

© 2014 Biophysical Society. Published by Elsevier Inc.

Additional details

Created:
August 19, 2023
Modified:
October 26, 2023