in situ rt-pcr optimized for electron microscopy allows description of subcellular morphology of target mrna-expressing cells in the brain

in situ rt-pcr optimized for electron microscopy allows description of subcellular morphology of target mrna-expressing cells in the brain

;Laura Cubas-Nuñez;María Duran-Moreno;Jessica Castillo-Villalba;Jorge Fuentes-Maestre;Bonaventura Casanova;José M. García-Verdugo;José M. García-Verdugo;Sara Gil-Perotín;Sara Gil-Perotín;Sara Gil-Perotín
macromolecular bioscience 2017 Vol. 11 pp. -
196
cubas-nuez2017frontiersin

Abstract

In situ RT-PCR detects and amplifies mRNA (cDNA) while obtaining spatial information of gene expression. When the intended use is an ultrastructural analysis of morphology, the procedure may be technically challenging and quality of tissue dramatically altered by proteolytic digestion and extreme astringency and temperature conditions. We describe a low-damaging protocol of in situ RT-PCR combined to conventional electron microscopy that preserves fine morphology, increases sensitivity, and decreases costs and complexity associated to RNA probes.

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Article ID:
150910
Unique Identifier:
10.3389/fncel.2017.00141
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