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Author Spotlight: Unlocking Insights into the Immune Cell Landscape of Tumors
Published on: August 18, 2023
Autostaining Immune Multiplex Immunohistochemistry Panels and Imaging Analysis
Hidetoshi Mori1,2, Alexander D Borowsky3
1Center for Immunology and Infectious Diseases, University of California, Davis, CA, USA. hmori@health.ucdavis.edu.
Insights
Multiplex immunohistochemistry visualizes human immune cells in tissues. This technique uses tyramide signal amplification for detailed cell profiling and localization, overcoming flow cytometry limitations.
Area of Science:
- Immunology
- Cell Biology
- Biotechnology
Background:
- Immune cell identification often requires multiple markers.
- Flow cytometry offers multi-parameter analysis but lacks spatial localization.
- Multiplex immunohistochemistry (IHC) enables simultaneous detection of multiple markers within the tissue microenvironment.
Purpose of the Study:
- To present standardized staining procedures for human immune-cell multiplex panels.
- To demonstrate the application of tyramide signal amplification-based IHC on an autostainer.
- To explore imaging analysis techniques for multiplex IHC data.
Main Methods:
- Development of multiplex IHC staining protocols for human immune cells.
- Utilized tyramide signal amplification (TSA) for enhanced signal detection.
- Employed automated staining on an autostainer for consistency.
- Applied advanced imaging analysis to interpret multiplex IHC results.
Main Results:
- Successfully established robust multiplex IHC staining procedures for human immune cells.
- Demonstrated the visualization of multiple immune cell markers on formalin-fixed paraffin-embedded (FFPE) tissues.
- Validated the utility of TSA-based IHC for high-sensitivity multiplexing.
- Identified effective imaging analysis strategies for multiplex IHC data.
Conclusions:
- Multiplex IHC is a powerful technique for immune cell profiling with spatial context.
- The presented protocols enable detailed characterization of immune cells within their native tissue environment.
- This approach complements flow cytometry by providing crucial localization information.
Abstract:
Immune cells often require several markers for positive identification. While flow-cytometry can provide detailed multi-parameter cell identification it does not retain the cell localization. Multiplex immunohistochemistry provides a valuable technique for cell profiling with localization in the tissue microenvironment. In this study, we present the staining procedures for human immune-cell multiplex panels, employing tyramide signal amplification-based immunohistochemistry on an autostainer. Additionally, we explore the imaging analysis techniques applied to the stained tissues. These panels enable the visualization of immune cell markers on formalin-fixed paraffin-embedded tissues.
