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Mouse spleen tissue as a staining intensity reference for immunohistochemistry
Yeonsook Moon1, Gyeongsin Park, Kyungja Han
1Department of Laboratory Medicine, Inha University Medical College, Incheon, Korea.
Insights
Mouse spleen tissue can serve as a reliable staining intensity reference for immunohistochemistry (IHC). This internal control enhances the reproducibility and accuracy of diagnostic and research IHC staining results.
Area of Science:
- Biomedical Sciences
- Pathology
- Immunohistochemistry
Background:
- Immunohistochemistry (IHC) is crucial for diagnostics and research.
- IHC's subjectivity and poor reproducibility limit its utility.
- A standardized internal reference is needed for accurate IHC interpretation.
Purpose of the Study:
- To evaluate the feasibility of using mouse spleen tissue as an internal staining intensity reference in conventional IHC.
- To assess the stability of mouse spleen staining under varying IHC procedural conditions.
Main Methods:
- Formalin-fixed, paraffin-embedded mouse spleen tissue was used.
- Tissue was subjected to variations in primary antibody types, antigen retrieval methods, chromogen exposure times, and secondary antibody concentrations.
- Mouse spleen was incorporated into tissue microarrays for comparison with the c-erbB2 IHC scoring system.
Main Results:
- Mouse spleen exhibited consistent staining intensity across different primary antibody types and minimal variation with antigen retrieval methods.
- Staining intensity in mouse spleen varied with chromogen exposure time and secondary antibody concentration.
- Splenic B cells and plasma cells in mouse spleen showed staining intensities comparable to scores 1+ and 3+ in c-erbB2 IHC, respectively.
Conclusions:
- Mouse spleen tissue demonstrates potential as a stable and reliable staining intensity reference for immunohistochemistry.
- Incorporating mouse spleen tissue can improve the objectivity and reproducibility of IHC interpretation.
- This method offers a practical solution to enhance the diagnostic accuracy of IHC assays.
Abstract:
Immunohistochemistry (IHC) is widely used in diagnostic practice and research, but it is limited due to its subjective nature and weakness in reproducibility. For successful interpretation, IHC requires an internal reference system that controls for procedural variables and provides a staining intensity reference. We investigated the feasibility of using mouse spleen tissue as an intensity reference in conventional IHC. Formalin-fixed, paraffin-embedded mouse (BALB/c) spleen tissue was stained with variable procedural conditions including primary antibody (Ab) types, antigen retrieval methods, chromogen exposure times, and secondary Ab concentrations. Mouse spleen tissue showed identical staining intensity regardless of primary Ab types, even without primary Ab, and showed minimal differences according to retrieval methods. However, it showed various staining intensities according to chromogen exposure time and secondary Ab concentration. When mouse spleen was included in tissue microarrays and compared with the c-erbB2 IHC scoring system, splenic B cells showed weak membrane staining compatible with score 1+, whereas splenic plasma cells showed strong staining intensity compatible with score 3+. These results show that mouse spleen tissue can serve as a staining intensity reference for the interpretation of IHC.
