Quantitative mRNA imaging throughout the entire Drosophila brain

Nat Methods. 2017 Jul;14(7):703-706. doi: 10.1038/nmeth.4309. Epub 2017 Jun 5.

Abstract

We describe a fluorescence in situ hybridization method that permits detection of the localization and abundance of single mRNAs (smFISH) in cleared whole-mount adult Drosophila brains. The approach is rapid and multiplexable and does not require molecular amplification; it allows facile quantification of mRNA expression with subcellular resolution on a standard confocal microscope. We further demonstrate single-mRNA detection across the entire brain using a custom Bessel beam structured illumination microscope (BB-SIM).

MeSH terms

  • Animals
  • Brain / metabolism*
  • Drosophila / anatomy & histology
  • Drosophila / metabolism*
  • In Situ Hybridization, Fluorescence / methods*
  • Microscopy / instrumentation
  • Microscopy / methods*
  • RNA, Messenger / metabolism*

Substances

  • RNA, Messenger