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Immunohistochemical Analysis in the Rat Central Nervous System and Peripheral Lymph Node Tissue Sections
Published on: November 14, 2016
Detecting proliferating cell nuclear antigen in archival rodent tissues
A Greenwell1, J F Foley, R R Maronpot
1National Toxicology Program, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709.
Environmental Health Perspectives
|December 1, 1993
Summary
This study introduces an enhanced immunohistochemical method for detecting proliferating cell nuclear antigen (PCNA) in archival rodent tissues. The improved technique allows sensitive detection of cell replication markers in long-term stored formalin-fixed tissues.
Area of Science:
- Histopathology
- Molecular Biology
- Toxicology
Background:
- Proliferating cell nuclear antigen (PCNA) is a key marker for cell replication.
- Detecting PCNA in archival formalin-fixed, paraffin-embedded tissues has been challenging due to reduced sensitivity.
- Existing methods are insufficient for reliable retrospective analysis of long-term stored samples.
Purpose of the Study:
- To develop and validate an enhanced immunohistochemical procedure for sensitive PCNA detection.
- To improve the detection of cell replication markers in aged, formalin-fixed rodent tissues.
- To enable retrospective analysis of archival tissues for toxicity and carcinogenicity studies.
Main Methods:
- An enhanced immunohistochemical technique involving microwave heating with heavy-metal salt antigen retrieval solution.
- Application of the method to archival rat and mouse tissues fixed in formalin.
- Evaluation of PCNA detection sensitivity in tissues stored for extended periods.
Main Results:
- Successfully applied an enhanced immunohistochemical procedure to detect PCNA in archival rodent tissues.
- Achieved consistent positive immunostaining for PCNA in formalin-fixed tissues (over 24 months) and paraffin blocks (up to 19 months).
- Demonstrated reliable detection of S-phase cells, indicating DNA replicative activity.
Conclusions:
- The enhanced method significantly improves PCNA detection sensitivity in challenging archival tissues.
- This technique facilitates retrospective analysis of previously stored rodent tissues.
- It is valuable for re-evaluating data from historical toxicity and carcinogenicity studies.
