Spatially Characterizing the Immune Contexture in Mouse Tissue Using Multiplex Immunohistochemistry

Lokman Pang; Matthias Ernst; Jennifer Huynh

doi:10.1007/978-1-0716-2811-9_20

Spatially Characterizing the Immune Contexture in Mouse Tissue Using Multiplex Immunohistochemistry

Methods Mol Biol. 2023:2593:307-316. doi: 10.1007/978-1-0716-2811-9_20.

Authors

Lokman Pang¹, Matthias Ernst¹, Jennifer Huynh²

Affiliations

¹ Olivia Newton-John Cancer Research Institute and School of Cancer Medicine, La Trobe University, Heidelberg, VIC, Australia.
² Olivia Newton-John Cancer Research Institute and School of Cancer Medicine, La Trobe University, Heidelberg, VIC, Australia. Jennifer.Huynh@onjcri.org.au.

PMID: 36513940
DOI: 10.1007/978-1-0716-2811-9_20

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.

Keywords: Cell surface markers; Immune cells; Multiplex immunohistochemistry; Spatial phenotyping; Tyramide signal amplification (TSA).

MeSH terms

Animals
Antibodies*
Biomarkers
Immunohistochemistry
Mice
Staining and Labeling

Substances

Antibodies
Biomarkers