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Fluorescence In Situ Hybridization with Quantum Dot Labels in E. coli Cells
Y. Liu, Z. Han, , A.M. Smith
Published in Humana Press Inc.
2021
PMID: 33576988
Volume: 2246
   
Pages: 141 - 155
Abstract
In this chapter we describe the use of fluorescent quantum dots (QDs) as labels for microbial mRNA transcripts using fluorescence in situ hybridization (FISH). Unlike organic dyes, which are the standard labels in modern FISH methods, QDs provide fluorescence signals that are much brighter and resistant to photobleaching, with an expanded spectral range for multiplexing. We describe the preparation of QDs with compact sizes necessary for accurate labeling, their application for analyzing lacZ transcripts in Escherichia coli cells using FISH, and an assessment of signal stability. We further discuss differences between methods for mammalian cells and bacteria, for which individual nucleic acids cannot be discretely counted due to the small cell size and the optical diffraction limit. © 2020, Springer Science+Business Media, LLC, part of Springer Nature.
About the journal
JournalMethods in Molecular Biology
PublisherHumana Press Inc.
ISSN10643745