Mouse spleen tissue as a staining intensity reference for immunohistochemistry.

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.