Imaging in cancer immunology: Phenotyping of multiple immune cell subsets in
situ in FFPE tissue sections

Short Communication - Immunology Case Reports (2021) Volume 4, Issue 1

immunobiology in recent years as a result of recent

successes in cancer immunotherapies and it is becoming

clear that immune cells play many sometimes conflicting

roles in the tumor microenvironment. However, obtaining

phenotypic information about the various immune cells that

play these roles in and around the tumor has been a

challenge. Existing methods can either deliver phenotypic

information on homogenous samples (e.g., flow cytometry

or PCR) or morphologic information on single

immunomarkers (standard IHC). We present here a

methodology for delivering quantitative per-cell marker

expression and phenotyping, analogous to that obtained

from flow cytometry but from cells imaged in situ in FFPE

tissue sections. This methodology combines the sequential

multi-marker labeling of up to 6 antigens using antibodies

all of the same species in a single section; automated

multispectral imaging (MSI) to remove the typically

problematic FFPE tissue auto fluorescence and correct

cross-talk between fluorescent channels and an automated

image analysis that can quantitate the per-cell marker

expression, determine the cellular phenotype, count these

cells separately in the tumor compartment and in the stroma

Author(s): Maciej P Zerkowski