A New Technology for Stabilization of Biomolecules in Tissues for Combined Histological and Molecular Analyses
- Autores: Christian Viertler, Daniel Groelz, Sibylle Gündisch, Karl Kashofer, Bilge Reischauer, Peter H. J. Riegman, Rosa Winther, Ralf Wyrich, Karl Friedrich Becker, Uwe Oelmüller, Kurt Zatloukal
- Localización: The Journal of molecular diagnostics, ISSN 1525-1578, Vol. 14, Nº. 5, 2012, págs. 458-466
- Idioma: inglés
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Resumen
For accurate diagnosis, prediction of outcome, and selection of appropriate therapies, the molecular characterization of human diseases requires analysis of a broad spectrum of altered biomolecules, in addition to morphological features, in affected tissues such as tumors. In a high-throughput screening approach, we have developed the PAXgene Tissue System as a novel tissue stabilization technology. Comprehensive characterization of this technology in stabilized and paraffin-embedded human tissues and comparison with snap-frozen tissues revealed excellent preservation of morphology and antigenicity, as well as outstanding integrity of nucleic acids (genomic DNA, miRNA, and mRNA) and phosphoproteins. Importantly, PAXgene-fixed, paraffin-embedded tissues provided RNA quantity and quality not only significantly better than that obtained with neutral buffered formalin, but also similar to that from snap-frozen tissue, which currently represents the gold standard for molecular analyses. The PAXgene tissue stabilization system thus opens new opportunities in a variety of molecular diagnostic and research applications in which the collection of snap-frozen tissue is not feasible for medical, logistic, or ethical reasons. Furthermore, this technology allows performing histopathological analyses together with molecular studies in a single sample, which markedly facilitates direct correlation of morphological disease phenotypes with alterations of nucleic acids and other biomolecules.
