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Updated: Jun 19, 2026
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Immunohistochemical Analysis in the Rat Central Nervous System and Peripheral Lymph Node Tissue Sections
Published on: November 14, 2016
Combined effects of formalin fixation and tissue processing on immunorecognition
D Otali1, C R Stockard, D K Oelschlager
1Department of Pathology, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA.
Summary
Tissue processing steps, including xylene exposure, significantly impact antigen detection in immunohistochemistry. Antigen retrieval helps but doesn't fully restore immunorecognition, highlighting the need for careful protocol optimization.
Area of Science:
- Cell Biology
- Immunohistochemistry
- Cancer Research
Background:
- Aldehyde-based fixation is known to affect antigen detection in immunodetection.
- The systematic effects of cumulative tissue processing steps on immunodetection remain underexplored.
Purpose of the Study:
- To investigate the impact of aldehyde fixation and sequential tissue processing steps on immunohistochemical antigen detection.
- To evaluate the role of antigen retrieval in mitigating these effects.
Main Methods:
- Cultured DU145 (prostate) and SKOV3 (ovarian) cancer cell lines on slides.
- Assessed immunohistochemical detection of Ki67/MIB-1 and PCNA after varying formalin fixation times and cumulative tissue processing steps.
- Evaluated the effect of antigen retrieval (AR) as a variable.
Main Results:
- Each tissue processing step, beyond fixation, influenced immunorecognition of target antigens.
- Xylene exposure caused the most significant reduction in immunorecognition.
- Antigen retrieval (AR) restored much of the staining but not all, and provided consistent results across processing steps.
Conclusions:
- Tissue processing steps critically affect immunohistochemical staining outcomes.
- Antigen retrieval is crucial for optimizing staining, particularly for PCNA, and should be considered for all steps.
- Cellular variations in antigen packaging may influence the susceptibility to fixation and processing effects.
