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Updated: May 8, 2026
Tumor Transplantation for Assessing the Dynamics of Tumor-Infiltrating CD8+ T Cells in Mice
Published on: June 12, 2021
Immunohistochemical assessment of immune cells in mouse tumor tissue
Lauren J Bayne1, Robert H Vonderheide
1Abramson Family Cancer Research Institute, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA.
Insights
This protocol details a histological method for evaluating immune cells while preserving tissue architecture. The technique involves cryosectioning, fixing, and antibody staining of frozen tissues for long-term storage and viewing.
Area of Science:
- Immunohistochemistry
- Tissue Histology
- Cell Biology
Background:
- Evaluating immune cell distribution and phenotype within intact tissue architecture is crucial for understanding immune responses.
- Traditional methods may compromise tissue integrity, limiting detailed histological analysis.
- A need exists for a reliable protocol that preserves tissue structure for immune cell evaluation.
Purpose of the Study:
- To describe a detailed protocol for the histological evaluation of immune cells.
- To present a method that preserves native tissue architecture.
- To enable long-term storage and viewing of stained tissue sections.
Main Methods:
- Utilizing frozen tissues embedded in optimal cutting temperature (OCT) medium immediately post-dissection.
- Cryosectioning tissues into thin (∼8 µm) sections and mounting onto microscope slides.
- Fixing sections (methanol or formaldehyde), followed by antibody staining for immune cell surface markers and detection via an avidin-biotin-based peroxidase system.
Main Results:
- Successful preservation of tissue architecture during the histological evaluation process.
- Specific immune cell populations identified through targeted antibody staining.
- Stained sections are stable for long-term storage and subsequent microscopic examination.
Conclusions:
- The described protocol provides a robust method for the histological assessment of immune cells.
- This technique allows for detailed analysis of immune cell localization within preserved tissue context.
- The protocol facilitates reproducible and durable immune cell profiling in histological samples.
Abstract:
This protocol describes a procedure for the evaluation of immune cells at the histological level. This technique preserves tissue architecture, making use of tissues that have been frozen in optimal cutting temperature (OCT) medium immediately after dissection and then cut with a cryostat into thin (∼8 µm) sections. The tissue slices are transferred to microscope slides and, as described here, are fixed with either methanol or formaldehyde. The fixed tissue sections are then stained with antibodies directed against cell surface markers expressed by the immune cells of interest. The antibodies are detected with an avidin-biotin-based peroxidase system, and, after mounting, the sections can be stored and viewed for years.
