Application of Immunohistochemical Staining to Detect Antigen Destruction as a Measure of Tissue Damage

Abdullah Onul, Michael D. Colvard, William A. Paradise, Kim M. Elseth, Benjamin J. Vesper, Eftychia Gouvas, Zane Deliu, Kelly D. Garcia, William J. Pestle, James A. Radosevich

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

Electrocautery and directed energy devices (DEDs) such as lasers, which are used in surgery, result in tissue damage that cannot be readily detected by traditional histological methods, such as hematoxylin and eosin staining. Alternative staining methods, including 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) to stain live tissue, have been reported. Despite providing superior detection of damaged tissue relative to the hematoxylin and eosin (H&E) method, the MTT method possesses a number of drawbacks, most notably that it must be carried out on live tissue samples. Herein, we report the development of a novel staining method, "antigen destruction immunohistochemistry" (ADI), which can be carried out on paraffin-embedded tissue. The ADI method takes advantage of epitope loss to define the area of tissue damage and provides many of the benefits of live tissue MTT staining without the drawbacks inherent to that method. In addition, the authors provide data to support the use of antibodies directed at a number of gene products for use in animal tissue for which there are no species-specific antibodies commercially available, as well as an example of a species-specific direct antibody. Data are provided that support the use of this method in many tissue models, as well as evidence that ADI is comparable to the live tissue MTT method.

Original languageEnglish (US)
Pages (from-to)683-693
Number of pages11
JournalJournal of Histochemistry and Cytochemistry
Volume60
Issue number9
DOIs
StatePublished - Sep 2012

Keywords

  • MTT
  • antigen destruction immunohistochemistry method
  • directed energy device
  • hematoxylin and eosin
  • immunohistochemistry
  • tissue damage/viability

ASJC Scopus subject areas

  • Anatomy
  • Histology

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  • Cite this

    Onul, A., Colvard, M. D., Paradise, W. A., Elseth, K. M., Vesper, B. J., Gouvas, E., Deliu, Z., Garcia, K. D., Pestle, W. J., & Radosevich, J. A. (2012). Application of Immunohistochemical Staining to Detect Antigen Destruction as a Measure of Tissue Damage. Journal of Histochemistry and Cytochemistry, 60(9), 683-693. https://doi.org/10.1369/0022155412452146