GCC Center for Advanced Microscopy and Image Informatics 1 article published in JoVE Biochemistry Single Cell Analysis Of Transcriptionally Active Alleles By Single Molecule FISH Ragini M. Mistry1,2, Pankaj K. Singh1,3, Maureen G. Mancini1,2, Fabio Stossi1,2, Michael A. Mancini1,2,3,4 1GCC Center for Advanced Microscopy and Image Informatics, 2Department of Molecular and Cellular Biology, Baylor College of Medicine, 3Center for Translational Cancer Research, Institute of Biosciences and Technology, Texas A&M University, 4Department of Pharmacology and Chemical Biology, Baylor College of Medicine Single molecule RNA fluorescence in situ hybridization (smFISH) is a method to accurately quantify levels and localization of specific RNAs at the single cell level. Here, we report our validated lab protocols for wet-bench processing, imaging and image analysis for single cell quantification of specific RNAs.