| Literature DB >> 32394381 |
Harry M T Choi1, Maayan Schwarzkopf1, Niles A Pierce2,3,4.
Abstract
In situ hybridization based on the mechanism of hybridization chain reaction (HCR) enables multiplexed quantitative mRNA imaging in the anatomical context of whole-mount vertebrate embryos. Third-generation in situ HCR (v3.0) provides automatic background suppression throughout the protocol, dramatically enhancing performance and ease of use. In situ HCR v3.0 supports two quantitative imaging modes: (1) qHCR imaging for analog mRNA relative quantitation with subcellular resolution in an anatomical context and (2) dHCR imaging for digital mRNA absolute quantitation with single-molecule resolution in an anatomical context. Here, we provide protocols for qHCR and dHCR imaging in whole-mount zebrafish, chicken, and mouse embryos.Entities:
Keywords: Accuracy; Autofluorescence; Automatic background suppression; Fluorescence in situ hybridization (FISH); Hybridization chain reaction (HCR); In situ HCR v3.0; Multiplexed; Precision; Quantitative; Signal amplification; Single-molecule imaging; Thick sample penetration; Whole-mount vertebrate embryos; dHCR imaging; mRNA imaging; qHCR imaging
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Year: 2020 PMID： 32394381 DOI： 10.1007/978-1-0716-0623-0_10
Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745