Application of immunohistochemical staining to detect antigen destruction as a measure of tissue damage.
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.
Center for Molecular Biology of Oral Diseases, College of Dentistry, University of Illinois at Chicago, Chicago, IL 60612, USA.
Abdullah
A
Onul
Michael D
MD
Colvard
William A
WA
Paradise
Kim M
KM
Elseth
Benjamin J
BJ
Vesper
Eftychia
E
Gouvas
Zane
Z
Deliu
Kelly D
KD
Garcia
William J
WJ
Pestle
James A
JA
Radosevich
D000818:Animals; D000906:Antibodies; D000918:Antibody Specificity; D000941:Antigens / Q000032:analysis* / Q000276:immunology; D004396:Coloring Agents; D003429:Cross Reactions; D004801:Eosine Yellowish-(YS); D005404:Fixatives; D005557:Formaldehyde; D006416:Hematoxylin; D006358:Hot Temperature; D007150:Immunohistochemistry / Q000379:methods*; D016612:Paraffin Embedding; D011489:Protein Denaturation; D017510:Protein Folding; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000032:analysis / Q000276:immunology; D013194:Staining and Labeling / Q000379:methods; D013552:Swine; D013778:Tetrazolium Salts; D013844:Thiazoles; D014443:Tyrosine / Q000031:analogs & derivatives / Q000032:analysis / Q000276:immunology
D000906:Antibodies; D000941:Antigens; D004396:Coloring Agents; D004801:Eosine Yellowish-(YS); D005404:Fixatives; D005557:Formaldehyde; D006416:Hematoxylin; D018719:Erb-b2 Receptor Tyrosine Kinases; D013778:Tetrazolium Salts; D013844:Thiazoles; D014443:Tyrosine; C002744:3-nitrotyrosine; C022616:thiazolyl blue