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

Nature Clinical Practice Urology volume 3, pages 268–278 (2006)Cite this article

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.

Key Points

  • Appropriate fixation techniques, employed soon after tissue sampling, are essential for the preservation of tissues

  • Specific chemical modifications occur in tissues during fixation and can interfere with subsequent analytical techniques, unless properly understood

  • All tissue fixation methods induce artefacts in preserved tissues, which can be physical (morphological) or chemical

  • Understanding the basis of the induced artefacts allows their presence to be compensated for, or even reversed, during subsequent analyses

  • Collection, preservation and storage of all human biological material are governed by law and it is imperative that appropriate consent and ethical approval are obtained before any study employing such material is undertaken

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Figure 1: Formaldehyde interacts with proteins by modifying lysine residues to generate intermolecular aldol cross-linkages.
Figure 2: Electropherograms and gel-like images of total RNA using the Agilent Bioanalyzer (Agilent).
Figure 3: Each strand of the 22 pairs of non-sex chromosomes (autosomes) comprises approximately 1.5 × 108 nucleotides and contains some 3 × 103 genes.

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Acknowledgements

The authors would like to thank Mrs Jill Gosney for editing the manuscript prior to submission.

Author information

Authors and Affiliations

  1. Professor and Director of Cellular Pathology and Molecular Genetics,

    Christopher S Foster, Christine M Gosden & Youqiang Q Ke

  2. Professor of Medical Genetics,

    Christopher S Foster, Christine M Gosden & Youqiang Q Ke

  3. Division of Pathology, Professor and Director of Molecular Pathology, School of Cancer Studies, University of Liverpool, Liverpool, UK

    Christopher S Foster, Christine M Gosden & Youqiang Q Ke

Authors
  1. Christopher S Foster
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  2. Christine M Gosden
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  3. Youqiang Q Ke
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Correspondence to Christopher S Foster.

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Foster, C., Gosden, C. & Ke, Y. Primer: tissue fixation and preservation for optimal molecular analysis of urologic tissues. Nat Rev Urol 3, 268–278 (2006). https://doi.org/10.1038/ncpuro0487

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