Cancer immunotherapy research has
increasingly leveraged knowledge about the
tumor microenvironment for the development
of novel therapeutics and targets. Advancing
our current understanding of the tumor
microenvironment will involve continued
characterization of the interactions that occur
among immune cells and cancer cells that
reside within the tumor and its periphery.
Fluorescent multiplex immunohistochemistry
(fIHC) assays are uniquely suited to
characterizing and quantifying these complex
interactions in situ. Here we describe a robust,
fully-automated 7-color fIHC method that
would significantly shorten the Opal™
procedure from days to overnight. We coupled
this automated staining procedure with a
multispectral imaging system for simultaneous
detection of up to 6 tissue biomarkers plus
nuclear counterstain, providing the ability to
visualize interactions between specific immune
and cancer cells within the context of the
tumor microenvironment.