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Multiplexed Imaging for Immune Profiling on Human FFPE Material.

Artur Mezheyeuski1, Carina Strell2

  • 1Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden. artur.mezheyeuski@igp.uu.se.

Methods in Molecular Biology (Clifton, N.J.)
|July 31, 2021
PubMed
Summary
This summary is machine-generated.

This chapter details a multiplex immunohistochemical staining pipeline for analyzing immune cells in human FFPE tissues. The method quantifies immune cell subtypes, their expression, and spatial distribution using multispectral imaging.

Keywords:
ImmunoprofilingMultiplex immunohistochemistryMultispectral imaging

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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.