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Related Experiment Video

Updated: Jun 29, 2025

Visualization, Quantification, and Mapping of Immune Cell Populations in the Tumor Microenvironment
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Imaging Mass Cytometry for In Situ Immune Profiling.

Kevin Hu1,2, Andrew Harman1,2, Heeva Baharlou3,4

  • 1Centre for Virus Research, The Westmead Institute for Medical Research, Westmead, NSW, Australia.

Methods in Molecular Biology (Clifton, N.J.)
|March 25, 2024
PubMed
Summary

This study presents an optimized protocol for imaging mass cytometry (IMC) combined with RNAscope, enabling simultaneous protein and RNA detection in tissues. This advanced technique enhances spatial resolution for immune profiling in various disease contexts.

Keywords:
Cell segmentationHuman tissue sectionsHyperion imaging systemImaging mass cytometryMetal conjugated antibodiesMultiplexed spatial mappingRNAscope detection

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Area of Science:

  • Biomedical Imaging
  • Molecular Pathology
  • Immunohistochemistry

Background:

  • Tissues exhibit complex cellular heterogeneity, requiring advanced imaging to resolve immune cell interactions.
  • Conventional methods like single-cell suspension lack spatial context crucial for understanding pathology.
  • High-dimensional imaging mass cytometry (IMC) allows simultaneous detection of up to 40 protein markers at subcellular resolution.

Purpose of the Study:

  • To optimize a staining protocol for imaging mass cytometry (IMC).
  • To integrate RNAscope technology with IMC for combined protein and RNA detection.
  • To expand IMC utility for investigating low-abundance or noncoding RNA targets within tissue microenvironments.

Main Methods:

  • Development and optimization of a novel staining protocol for IMC.
  • Integration of RNAscope in situ hybridization with IMC.
  • Validation of the protocol for concurrent protein and single-molecule RNA detection in tissue samples.

Main Results:

  • Successful integration of RNAscope with IMC, enabling multiplexed protein and RNA analysis.
  • Demonstration of enhanced capability for detecting low-abundance and noncoding RNA targets.
  • Preservation of subcellular resolution for both protein and RNA markers.

Conclusions:

  • The optimized protocol significantly expands the capabilities of IMC for comprehensive immune profiling.
  • This method provides crucial spatial context for studying host-pathogen interactions, tumor microenvironments, and inflammatory diseases.
  • The combined approach offers a powerful tool for advancing our understanding of complex biological systems within their native tissue context.