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Single Cell Analysis Of Transcriptionally Active Alleles By Single Molecule FISH
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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
Chapters
- 00:04Introduction
- 00:34RNA Fluorescence In Situ Hybridization (FISH) Fixation
- 01:14RNA FISH Hybridization
- 02:16Imaging Preparation
- 03:08High Resolution Microscopy and Image Processing
- 04:05Image Analysis
- 04:45Results: Representative Image Segmentation and E2-Induced GREB1 Quantification
- 05:56Conclusion
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.
