A Hitchhiker's guide to high-dimensional tissue imaging with multiplexed ion beam imaging.

Methods in cell biology
Authors
Keywords
Abstract

Advancements in multiplexed tissue imaging technologies are vital in shaping our understanding of tissue microenvironmental influences in disease contexts. These technologies now allow us to relate the phenotype of individual cells to their higher-order roles in tissue organization and function. Multiplexed Ion Beam Imaging (MIBI) is one of such technologies, which uses metal isotope-labeled antibodies and secondary ion mass spectrometry (SIMS) to image more than 40 protein markers simultaneously within a single tissue section. Here, we describe an optimized MIBI workflow for high-plex analysis of Formalin-Fixed Paraffin-Embedded (FFPE) tissues following antigen retrieval, metal isotope-conjugated antibody staining, imaging using the MIBI instrument, and subsequent data processing and analysis. While this workflow is focused on imaging human FFPE samples using the MIBI, this workflow can be easily extended to model systems, biological questions, and multiplexed imaging modalities.

Year of Publication
2024
Journal
Methods in cell biology
Volume
186
Pages
213-231
Date Published
12/2024
ISSN
0091-679X
DOI
10.1016/bs.mcb.2024.02.018
PubMed ID
38705600
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