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Hybridization Chain Reaction RNA Whole-Mount Fluorescence In situ Hybridization of Chemosensory Genes in Mosquito Olfactory Appendages
Published on: November 17, 2023
Methodological framework for chromogenic mRNA detection using in situ hybridization chain reaction.
Mitsuru Yashiro1,2, Yousuke Tsuneoka3, Yusuke Atsumi1,2
1Department of Anatomy, Faculty of Medicine, Toho University, Tokyo, Japan.
Histochemistry and Cell Biology
|May 6, 2026
Summary
A new chromogenic in situ hybridization chain reaction (HCR) method allows sensitive visualization of single mRNA molecules. This bright-field technique overcomes limitations of fluorescence HCR in routine pathology and autofluorescent tissues.
Area of Science:
- Molecular Biology
- Histology
- Biochemistry
Background:
- In situ hybridization chain reaction (HCR) is a sensitive single-molecule mRNA detection method.
- Conventional fluorescence HCR is limited in routine pathology and autofluorescent tissues.
Purpose of the Study:
- To develop a chromogenic in situ HCR protocol for sensitive mRNA detection.
- To expand HCR utility in histology and pathology.
Main Methods:
- Established a chromogenic in situ HCR protocol using short hairpin DNA and indirect hapten labeling.
- Validated the protocol on mouse tissues (brain, kidney, liver) and different section types.
- Demonstrated duplex staining and combined mRNA-protein visualization.
Main Results:
- Achieved sensitivity comparable to fluorescent HCR.
- Enabled clear visualization of low-abundance transcripts (Oxtr, Esr1).
- Showcased versatility in multiplexing and combining with immunohistochemistry.
Conclusions:
- Chromogenic in situ HCR offers high sensitivity with practical bright-field detection advantages.
- This method enhances HCR applicability in histological research and routine diagnostics.
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A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...
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