Fly (Dec 2025)

A fast in situ hybridization chain reaction method in Drosophila embryos and ovaries

  • Kyohei Mikami,
  • Yasuhiro Kozono,
  • Masaki Masukawa,
  • Satoru Kobayashi

DOI
https://doi.org/10.1080/19336934.2024.2428499
Journal volume & issue
Vol. 19, no. 1

Abstract

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The in situ hybridization chain reaction (isHCR) is a powerful method for visualizing mRNA in many species. We present a rapid isHCR method for Drosophila embryos and ovaries. Ethylene carbonate was added to the hybridization buffer to facilitate the hybridization reaction, and a modified short hairpin DNA was used in the amplification reaction; these modifications decreased the RNA staining time from 3 days to 1 day. This method is compatible with immunohistochemistry and can detect multiple mRNAs. The proposed method could significantly reduce staining time for Drosophila researchers using isHCR.

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