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Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging
Published on: April 8, 2016
13.5K
Systems pathology by multiplexed immunohistochemistry and whole-slide digital image analysis
Sami Blom1, Lassi Paavolainen1, Dmitrii Bychkov1
1Institute for Molecular Medicine Finland FIMM, University of Helsinki, Helsinki, Finland.
Scientific Reports
|November 16, 2017
Summary
A new multiplexed immunohistochemistry (mIHC) platform enables high-resolution, whole-slide imaging and analysis of up to six protein markers in tissues. This robust tool allows for quantitative, spatially resolved tissue cytometry in formalin-fixed human tumors.
Area of Science:
- Pathology
- Biotechnology
- Computational Biology
Background:
- Molecular histopathology is transitioning from single-marker immunohistochemistry (IHC) to multiplexed detection for understanding complex disease processes.
- Current systems lack the capability for high-resolution, multiplexed IHC (mIHC) with whole-slide imaging and analysis suitable for routine use.
Purpose of the Study:
- To develop and validate a novel mIHC platform for high-throughput, multiplexed biomarker detection and analysis in formalin-fixed tissues.
- To enable simultaneous detection and quantification of multiple protein markers and cell types at a cellular level within intact tissue architecture.
Main Methods:
- Development of an mIHC platform integrating fluorescent and chromogenic staining with automated whole-slide imaging.
- Implementation of integrated image analysis software for automatic quantification and classification of cells and tissue structures.
- Application of the platform in two proof-of-concept studies on human prostate cancer tissues.
Main Results:
- Simultaneous detection of six protein markers and nuclei in formalin-fixed paraffin-embedded tissues.
- Automated quantification and classification of over 128,000 immune cells and analysis of T cell subpopulations in tumor compartments.
- Automatic classification of over 83,000 epithelial cells and glands, with simultaneous analysis of androgen receptor (AR) and alpha-methylacyl-CoA (AMACR) expression.
Conclusions:
- The open-source 8-plex mIHC platform combined with whole-slide imaging and analysis offers a robust tool for quantitative, spatially resolved tissue cytometry.
- This technology facilitates improved characterization of histology and the tumor microenvironment directly in human tumor tissues.
- The platform supports the paradigm shift towards multiplexed biomarker analysis in routine histopathology for enhanced diagnostic and research capabilities.
