Assessing the Practical Application of Imaging Mass Cytometry for Visualizing Tumor Immune Microenvironment Heterogeneity

Shengtao Huang, Tianhao Zhou, Shaoqian Du, Shanhe Li, Ning Li, Ziqi Xv, Mengyi Jiang, Ying Zhu, Chaoyong Yang, Hongxia Wang, Xianting Ding, Lei Tang, Lijuan Zang

Article ID: 7865
Vol 38, Issue 2, 2024
DOI: https://doi.org/10.23812/j.biol.regul.homeost.agents.20243802.134
Received: 20 February 2024; Accepted: 20 February 2024; Available online: 20 February 2024; Issue release: 20 February 2024

Abstract

Background: Analyses of immune cell subsets and immune indices are becoming increasingly important. By interacting molecular probe and immunohistochemistry (IHC) analysis with laser ablation, imaging mass cytometry (IMC) offers high-dimensional in situ measurements in tissue slides with a spatial resolution of 1 μm. One of the most significant challenges is to implement stringent quality control strategies during the application of IMC to facilitate reproducible data analysis. We have developed a comprehensive protocol for clinical and experimental use by comparing IMC to standard clinical immunohistochemical approaches. Methods: We discussed the multi-step experimental processing procedures of IMC, including specimen preparation, panel design, antibody selection, lanthanide metal labelling, and data pre-processing workflows by IHC and fluorescent multiplex immunohistochemistry (mIHC) analysis. Based on IMC, we developed a standard operation and a well-established agreement for 29 breast cancer (BC) patients to identify a structured tumor immune microenvironment. Results: Metal labelling has no effect on the specificity of the antibodies target (p > 0.05). The 3 μm-thick formalin-fixed and paraffin-embedded (FFPE) and fresh-frozen slides exhibited the least blur (p < 0.01) and the lowest nonspecific adsorption of antibodies (p < 0.01). A detailed analysis of human breast cancer tissues revealed a wide range of differences in the composition and frequency of immune cells within the tumor microenvironment. Conclusions: This study presents optimized proposals on procedures for IMC analysis and a standardized quality control strategy.


Keywords

imaging mass cytometry (IMC);Cytometry by Time-Of-Flight (CyTOF);quality control;tumor microenvironment;breast cancer


References

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