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, multiplexable and does not require molecular amplification; it allows facile mRNA expression quantification with subcellular resolution on a standard confocal microscope. Using a custom Bessel Beam-Structured Illumination microscope (BB-SIM), we further demonstrate single-mRNA detection across the entire brain sample.
Footnotes
4 Department of Anatomy and Structural Biology, Dominick P. Purpura Department of Neuroscience, Gruss Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx, NY, 10461;
5 Institute for Systems Genetics, Department of Cell Biology, New York University Langone Medical Center, New York, NY