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In situ hybridization methods are used across the biological sciences to map mRNA expression within intact specimens. Multiplexed experiments, in which multiple target mRNAs are mapped in a single sample, are essential for studying regulatory interactions, but remain cumbersome in most model organisms. Programmable in situ amplifiers based on the mechanism of hybridization chain reaction (HCR) overcome this longstanding challenge by operating independently within a sample, enabling multiplexed experiments to be performed with an experimental timeline independent of the number of target mRNAs. To assist biologists working across a broad spectrum of organisms, we demonstrate multiplexed in situ HCR in diverse imaging settings: bacteria, whole-mount nematode larvae, whole-mount fruit fly embryos, whole-mount sea urchin embryos, whole-mount zebrafish larvae, whole-mount chicken embryos, whole-mount mouse embryos and formalin-fixed paraffin-embedded human tissue sections. In addition to straightforward multiplexing, in situ HCR enables deep sample penetration, high contrast and subcellular resolution, providing an incisive tool for the study of interlaced and overlapping expression patterns, with implications for research communities across the biological sciences.

Original publication

DOI

10.1242/dev.140137

Type

Journal article

Journal

Development

Publication Date

01/10/2016

Volume

143

Pages

3632 - 3637

Keywords

Bacteria, Whole-mount embryos and larvae, Deep sample penetration, High contrast, Hybridization chain reaction (HCR), In situ amplification, In situ hybridization, Multiplexing, Subcellular resolution, Tissue sections, Animals, Drosophila, Embryo, Nonmammalian, Humans, In Situ Hybridization, RNA, Messenger, Zebrafish