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Non‐radioactive in situ detection of mRNA in ES cell‐derived cardiomyocytes and in the developing heart
Author(s) -
Fijnvandraat Arnoud C.,
De Boer Piet A.J.,
Deprez Ronald H. Lekanne,
Moorman Antoon F.M.
Publication year - 2002
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.10154
Subject(s) - in situ , microbiology and biotechnology , embryonic stem cell , biology , in situ hybridization , gene expression , in situ hybridisation , messenger rna , context (archaeology) , gene , cell , chemistry , genetics , paleontology , organic chemistry
Non‐radioactive in situ hybridisation is an excellent method to visualise mRNA molecules within their topographical context. Recently we have reported a new non‐radioactive in situ hybridisation procedure on tissue sections that is essentially based on the whole mount in situ hybridisation procedure. This method is superior in spatial resolution and sensitivity compared to the radioactive in situ hybridisation procedure. Generally, low levels of gene expression, such as found with the developmental onset of gene expression and in differentiating embryonic stem cells, are difficult to detect by in situ hybridisation. Here an application of the protocol is presented which is based on tyramide signal amplification, which enables the detection of very low abundant mRNAs. The significance of this method is two‐fold: (1) the molecular phenotype of embryonic stem cell‐derived cardiomyocytes can be examined at the cellular level with high sensitivity, and (2) the number of cells that express the gene of interest can be assessed. Microsc. Res. Tech. 58:387–394, 2002. © 2002 Wiley‐Liss, Inc.