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Single Cell Analysis Of Transcriptionally Active Alleles By Single Molecule FISH
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.
