Protocol for multispectral imaging on cryosections to map myeloid cell heterogeneity in its spatial context

STAR Protocols, vol. 4, pp. 102601

Abstract

Recent technical advances, such as single-cell RNA sequencing and mass cytometry, improve identification of cell types and subsets in a range of healthy and diseased tissues at the expense of their cellular and molecular context. Here, we present a protocol for in situ multispectral imaging to map myeloid cell heterogeneity in tissue cryosections, describing steps for cutting sequential sections, antibody titration, and building a spectral library. We then detail procedures for multispectral imaging and preparing data for downstream analysis. For complete details on the use and execution of this protocol, please refer to Goossens et al. (2022).1

Authors 6

1. Elias B Wieland

   Maastricht University Medical Centre · Maastricht University

   Affiliation as printed

   Cardiovascular Research Institute Maastricht, Experimental Vascular Pathology, Department of Pathology, Maastricht University Medical Center+, Maastricht, The Netherlands

2. Laura JAP Kempen

   University of Liège · Maastricht University Medical Centre · Maastricht University

   Affiliation as printed

   Cardiovascular Research Institute Maastricht, Experimental Vascular Pathology, Department of Pathology, Maastricht University Medical Center+, Maastricht, The Netherlands; Laboratory of Immunology and Vaccinology, Faculty of Veterinary Medicine, FARAH, Liège University, Liège, Belgium; Laboratory of Immunophysiology, GIGA Institute, Liège University, Liège, Belgium

   Cardiovascular Research Institute Maastricht, Experimental Vascular Pathology, Department of Pathology, Maastricht University Medical Center+, Maastricht, The Netherlands

   Laboratory of Immunology and Vaccinology, Faculty of Veterinary Medicine, FARAH, Liège University, Liège, Belgium

   Laboratory of Immunophysiology, GIGA Institute, Liège University, Liège, Belgium

3. Chang Wei Lu

   Maastricht University Medical Centre · Maastricht University

   Affiliation as printed

   Cardiovascular Research Institute Maastricht, Experimental Vascular Pathology, Department of Pathology, Maastricht University Medical Center+, Maastricht, The Netherlands; Institute for Computational Biomedicine, Heidelberg, Germany

   Cardiovascular Research Institute Maastricht, Experimental Vascular Pathology, Department of Pathology, Maastricht University Medical Center+, Maastricht, The Netherlands

   Institute for Computational Biomedicine, Heidelberg, Germany

4. Marjo M. P. C. Donners

   Maastricht University Medical Centre · Maastricht University

   Affiliation as printed

   Cardiovascular Research Institute Maastricht, Experimental Vascular Pathology, Department of Pathology, Maastricht University Medical Center+, Maastricht, The Netherlands

5. Erik A.L. Biessen Aachen Institute for Molecular Cardiovascular Research (IMCAR)

   Maastricht University Medical Centre · Maastricht University · RWTH Aachen University

   Affiliation as printed

   Cardiovascular Research Institute Maastricht, Experimental Vascular Pathology, Department of Pathology, Maastricht University Medical Center+, Maastricht, The Netherlands; Institute for Molecular Cardiovascular Research, RWTH Aachen University, Aachen, Germany

   Cardiovascular Research Institute Maastricht, Experimental Vascular Pathology, Department of Pathology, Maastricht University Medical Center+, Maastricht, The Netherlands

   Institute for Molecular Cardiovascular Research, RWTH Aachen University, Aachen, Germany

6. Pieter Goossens corresponding

   Maastricht University Medical Centre · Maastricht University

   Affiliation as printed

   Cardiovascular Research Institute Maastricht, Experimental Vascular Pathology, Department of Pathology, Maastricht University Medical Center+, Maastricht, The Netherlands. Electronic address: pieter.goossens@maastrichtuniversity.nl

   Cardiovascular Research Institute Maastricht, Experimental Vascular Pathology, Department of Pathology, Maastricht University Medical Center+, Maastricht, The Netherlands