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Updated: May 21, 2026
Histological-Based Stainings Using Free-Floating Tissue Sections
Published on: August 25, 2020
Application of immunohistochemical staining to detect antigen destruction as a measure of tissue damage
Abdullah Onul1, Michael D Colvard, William A Paradise
1Center for Molecular Biology of Oral Diseases, College of Dentistry, University of Illinois at Chicago, Chicago, IL 60612, USA.
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.
Insights
A novel "antigen destruction immunohistochemistry" (ADI) staining method detects surgical tissue damage in paraffin-embedded samples. This method offers benefits similar to live tissue staining without its drawbacks, improving histological analysis.
Area of Science:
- Histopathology
- Surgical Pathology
- Immunohistochemistry
Background:
- Traditional histological methods like H&E staining fail to detect tissue damage from electrocautery and directed energy devices (DEDs).
- While MTT staining can detect live tissue damage, it requires live samples, limiting its application.
- There is a need for reliable methods to assess DED-induced tissue damage in preserved samples.
Purpose of the Study:
- To develop a novel staining method for detecting tissue damage caused by surgical energy devices.
- To create a method applicable to paraffin-embedded tissues, overcoming limitations of live-tissue staining.
- To validate the efficacy of the new method using various tissue models and antibody applications.
Main Methods:
- Development of "antigen destruction immunohistochemistry" (ADI), a novel staining technique.
- Utilizing epitope loss in paraffin-embedded tissues to identify areas of damage.
- Employing antibodies for gene products, including those for which species-specific antibodies are not commercially available.
Main Results:
- The ADI method successfully identifies tissue damage in paraffin-embedded samples.
- ADI provides benefits comparable to live tissue MTT staining but without the requirement for live samples.
- The study demonstrates the utility of ADI across multiple tissue models and antibody types, including species-specific antibodies.
Conclusions:
- ADI is a novel and effective method for detecting surgical DED-induced tissue damage in paraffin-embedded tissues.
- This technique offers a valuable alternative to existing methods, overcoming the limitations of MTT staining.
- ADI enhances the ability to analyze tissue damage in surgical pathology research and practice.
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