Journal
DEVELOPMENT
Volume 148, Issue 22, Pages -Publisher
COMPANY BIOLOGISTS LTD
DOI: 10.1242/dev.199847
Keywords
Immunofluorescence (IF); RNA fluorescence in situ hybridization (RNA-FISH); qHCR imaging; Formalin-fixed paraffin-embedded (FFPE) mouse brain and human breast tissue sections; Whole-mount zebrafish embryos
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Funding
- National Institutes of Health [NIBIB R01EB006192, NIGMS R44GM140796]
- Defense Advanced Research Projects Agency [HR0011-17-2-0008]
- Beckman Institute, California Institute of Technology (Programmable Molecular Technology Center, PMTC)
- California Institute of Technology
- National Institutes of Health (NIGMS) [GM008042]
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This technology utilizes the HCR mechanism to achieve multiplexed, quantitative, high-resolution RNA imaging in highly autofluorescent samples, while also enabling accurate and precise protein relative quantitation in immunohistochemistry.
RNA in situ hybridization based on the mechanism of the hybridization chain reaction (HCR) enablesmultiplexed, quantitative, high-resolution RNA imaging in highly autofluorescent samples, including whole-mount vertebrate embryos, thick brain slices and formalin-fixed paraffin-embedded tissue sections. Here, we extend the benefits of one-step, multiplexed, quantitative, isothermal, enzyme-free HCR signal amplification to immunohistochemistry, enabling accurate and precise protein relative quantitation with subcellular resolution in an anatomical context. Moreover, we provide a unified framework for simultaneous quantitative protein and RNA imaging with one-step HCR signal amplification performed for all target proteins and RNAs simultaneously.
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