Primer: tissue fixation and preservation for optimal molecular analysis of urologic tissues

Abstract

Appreciation of the different methods of tissue handling is a prerequisite to obtaining accurate and biologically relevant tissue-based information. When a tissue sample is removed from its environment, biological changes are induced within its constituent cell population. It is inevitable that artefacts will be induced through obtaining and processing tissues, irrespective of whether the samples comprise a few cells derived by fine-needle aspiration or larger specimens obtained surgically. Depending upon the level of sophistication of the analytical methods subsequently employed, such changes might be irrelevant, or might result in acquisition of spurious data. While even brief ischemia alters expression of some genes, detectable by appropriate molecular techniques, the same changes might make no appreciable difference to tissue histomorphology. Furthermore, the phenotype of viable cells is known to change during tissue collection and handling. For example, transitional epithelial cells voided in urine are not phenotypically identical to those retained within the urothelium. Such phenotypic changes are temporary and might be of little consequence to subsequent analyses. Surprisingly, many cells in tissues preserved in an ischemic state can remain viable for several hours, and are believed to remain genotypically stable in the short term.