Multiplexed Imaging for Immune Profiling on Human FFPE Material

Area of Science:

• Immunohistochemistry
• Cell Biology
• Tissue Imaging

Background:

• Multiplex immunohistochemistry (IHC) enables simultaneous detection of multiple targets within a single tissue sample.
• Formalin-fixed paraffin-embedded (FFPE) tissues are a standard resource in biomedical research, but preserving antigenicity for multiplex IHC can be challenging.
• Understanding the spatial relationships and co-expression of immune cells is crucial for deciphering tissue microenvironments.

Purpose of the Study:

• To present a comprehensive pipeline for multiplex immunohistochemical staining, multispectral image acquisition, and subsequent analysis.
• To adapt and optimize the protocol for human formalin-fixed paraffin-embedded (FFPE) tissues.
• To enable quantitative assessment of immune cell subtypes, including dendritic cells, myeloid cells, and macrophages, alongside cytokeratin expression.

Main Methods:

• Multiplex immunohistochemical staining protocol optimized for FFPE human tissues.
• Multispectral imaging system for high-resolution image acquisition.
• Image analysis software for quantitative assessment of marker co-expression and spatial localization.

Main Results:

• Successful implementation of a multiplex IHC pipeline on human FFPE tissues.
• Quantitative data on the expression levels of dendritic cells, myeloid cells, macrophages, and cytokeratin.
• Detailed spatial localization mapping of immune cell subtypes within the tissue context.

Conclusions:

• The described pipeline provides a robust method for analyzing immune cell populations in FFPE tissues.
• This approach facilitates quantitative evaluation of immune cell (co-)expression and spatial distribution.
• The protocol is valuable for research involving tissue microenvironment analysis and immune cell profiling.