Simultaneous Detection of RNAs and Proteins with Subcellular Resolution
| Autor: | Koei Chin, Sunjong Kwon, Michel Nederlof |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Resolution (mass spectrometry) medicine.diagnostic_test Chemistry RNA Subcellular localization Molecular biology Staining 03 medical and health sciences chemistry.chemical_compound 030104 developmental biology 0302 clinical medicine 030220 oncology & carcinogenesis BDNA test Fluorescence microscope medicine DNA Fluorescence in situ hybridization |
| Zdroj: | Methods in Molecular Biology ISBN: 9781071606797 |
| Popis: | We describe the detailed methods of "immunoFISH" to analyze the expression level and the spatial localization of RNA transcripts and proteins on cultured cells and formal-fixed, paraffin-embedded (FFPE) tissue sections. On cultured cells, we detect specific transcripts using the Stellaris fluorescence in situ hybridization (FISH) probes labeled with fluorophores that target multiple regions along the desired transcripts and proteins combining the immunofluorescent staining. On FFPE tissue sections, we use the RNAscope FISH probes, modified branched DNA (bDNA) probes to amplify the RNA signals, followed by immunofluorescent staining for protein detection. The abundance, composition, and spatial distribution are determined by signals from fluorescently labeled proteins and individual transcripts of images acquired using high-resolution fluorescence microscopy. |
| Databáze: | OpenAIRE |
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