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Standardization in Immunohistology

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Formalin-Fixed Paraffin-Embedded Tissues

Part of the book series: Methods in Molecular Biology ((MIMB,volume 724))

Abstract

The rapid acceptance of immunohistology as an invaluable adjunct to morphologic diagnosis has been possible because of the development of new and more sensitive antibodies and detection systems that allow its application to formalin-fixed, paraffin-embedded tissue (FFPT). More importantly, antigen-retrieval techniques have resulted in some degree of consistency allowing immunohistology to be used reliably as a diagnostic tool. The advent of prognostic and predictive biomarkers, and the desire for individualized therapy has resulted in mounting pressure to employ the immunohistological assay in a quantitative manner. While it was not a major issue when the technique was employed in a qualitative manner, the numerous variables in the preanalytical and analytical phases of the test procedure that influence the immunoexpression of proteins in FFPT become critical to standardization. Tissue fixation is pivotal to antigen preservation but exposure to fixative prior to accessioning by the laboratory is not controlled. Antigen retrieval, crucial in the analytical phase, continues to be employed in an empirical manner with the actual mechanism of action remaining elusive. There is great variation in reagents, methodology, and duration of tissue processing and immunostaining procedure, and the detection systems employed are not standardized between laboratories. While many of these variables are offset by the application of antigen retrieval, which enables the detection of a wide range of antigens in FFPT, the method itself is not standardized. This myriad of variables makes it inappropriate to provide meaningful comparisons of results obtained in different laboratories and even in the same laboratory, as in current practice, each specimen experiences different preanalytical variables. Furthermore, variables in interpretation exist and cutoff thresholds for positivity differ. Failure to recognize false-positive and false-negative stains leads to further errors of quantitative measurement. Many of the problems relating to the technology and interpretation of immunostaining originate from failure to recognize that this procedure is different from other histological stains and involves many more steps that cannot be monitored until the end result is attained. While several remedial measures can be suggested to address some of these problems, accurate and reproducible quantitative assessment of immunostains presently remains elusive as important variables that impact on antigen preservation in the paraffin-embedded biopsy ­cannot be standardized.

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Authors and Affiliations

  1. University of Newcastle, Newcastle, NSW, Australia

    Anthony S.-Y. Leong

  2. Peking University, Beijing, China

    Anthony S.-Y. Leong

  3. Victorian Cytology Service, Melbourne, VIC, Australia

    Trishe Y.-M. Leong

Authors
  1. Anthony S.-Y. Leong
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  2. Trishe Y.-M. Leong
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Correspondence to Anthony S.-Y. Leong .

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Editors and Affiliations

  1. Health Science Center, Dept. of Pathology, Kuwait University, Jabriya, Block 3B, The Forth Ring Road, Safat, Kuwait

    Fahd Al-Mulla

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Leong, A.SY., Leong, T.YM. (2011). Standardization in Immunohistology. In: Al-Mulla, F. (eds) Formalin-Fixed Paraffin-Embedded Tissues. Methods in Molecular Biology, vol 724. Humana Press. https://doi.org/10.1007/978-1-61779-055-3_3

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  • DOI: https://doi.org/10.1007/978-1-61779-055-3_3

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-054-6

  • Online ISBN: 978-1-61779-055-3

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