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Spatially Characterizing the Immune Contexture in Mouse Tissue Using Multiplex Immunohistochemistry
Lokman Pang1, Matthias Ernst1, Jennifer Huynh2
1Olivia Newton-John Cancer Research Institute and School of Cancer Medicine, La Trobe University, Heidelberg, VIC, Australia.
Insights
Multiplex immunohistochemistry (mIHC) allows simultaneous detection of multiple immune cell markers on one tissue slide. This protocol details mIHC for identifying T and B cells in mouse tissues, adaptable for other immune cell markers.
Area of Science:
- Immunology
- Histology
- Biotechnology
Background:
- Multiplex immunohistochemistry (mIHC) enables simultaneous detection of multiple biomarkers on a single tissue section.
- Characterizing immune cell populations is crucial for understanding tissue microenvironments.
Purpose of the Study:
- To describe a detailed protocol for mIHC staining.
- To identify distinct lymphocyte populations (T and B cells) in murine tissues.
Main Methods:
- Developed an mIHC staining workflow.
- Utilized specific antibodies targeting CD4, CD8α, FOXP3, and B220.
- Applied the protocol to murine tissue sections.
Main Results:
- Successfully identified distinct T cell (CD4+, CD8α+, FOXP3+) and B cell (B220+) populations.
- Demonstrated the feasibility of multiplex staining for immune cell profiling.
Conclusions:
- The described mIHC protocol is effective for simultaneous identification of key lymphocyte subsets.
- This strategy is adaptable for evaluating diverse immune cell markers and immune contexture in various murine tissues.
Abstract:
Multiplex immunohistochemistry (mIHC) facilitates the simultaneous detection of various immune cell markers on a single tissue section. Here, we describe a protocol for an mIHC staining workflow using specific antibodies against CD4, CD8α, FOXP3, and B220 to identify distinct lymphocyte populations including T and B cells. This staining strategy can be adapted to include other cell markers to evaluate the immune contexture in murine tissues.
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