Protocol for open-source automated universal high-content multiplex fluorescence for RNA in situ analysis

Autor: Jean Descarpentrie, Florian Bernard, Wilfried Souleyreau, Lucie Brisson, Thomas Mathivet, Ioannis S. Pateras, Océane C.B. Martin, Maria Lopez Chiloeches, Teresa Frisan
Jazyk: angličtina
Rok vydání: 2024
Předmět:
Zdroj: STAR Protocols, Vol 5, Iss 4, Pp 103508- (2024)
Druh dokumentu: article
ISSN: 2666-1667
DOI: 10.1016/j.xpro.2024.103508
Popis: Summary: In situ hybridization visualizes RNA in cells, but image analysis is complex. We present a protocol based on open-source software for automated high-content multiplex fluorescence in situ transcriptomics analysis. Steps include nuclei segmentation with a Fiji macro and quantification of up to 14 mRNA probes per image. We describe procedures for storing raw data, quality control images, and the use of a Python app to summarize all the results in one spreadsheet detailing the number of single or co-positive cells. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
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