Application of Automated mRNA In Situ Hybridization for Formalin-Fixed, Paraffin-Embedded Mouse Skin Sections

    June 2003 in “ Applied immunohistochemistry & molecular morphology ”
    Hiroaki Nitta, Jiro Kishimoto, Thomas M. Grogan
    TLDR Combining cell conditioning with mild protease digestion effectively shows versican mRNA in mouse skin sections.
    The study introduced an automated mRNA in situ hybridization application for the Ventana Discovery instrument, aimed at visualizing versican expression in formalin-fixed mouse skin paraffin sections. The research focused on the effects of protease digestion and heating pretreatment, known as cell conditioning, on hybridization signal detection. It was found that cell conditioning alone did not produce a signal, while the highest strength of protease digestion resulted in background staining. However, combining cell conditioning with mild protease digestion effectively demonstrated the versican mRNA signal in the hair papilla region. This indicated that the combination of these pretreatments enhanced mRNA target staining compared to using either method alone. The study confirmed that the automated process was applicable, simple, and reproducible for formalin-fixed mouse skin sections, with precise control over temperature and enzyme dosage to optimize assay conditions.
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