Antibody; Immunology; Microscopy; Molecular Biology; Single Cell; Biochemistry, Genetics and Molecular Biology (all); Immunology and Microbiology (all); General Immunology and Microbiology; General Biochemistry, Genetics and Molecular Biology; multispectral; imaging; multiplex
Abstract :
[en] 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).
Wieland, Elias B ✱; Cardiovascular Research Institute Maastricht, Experimental Vascular Pathology, Department of Pathology, Maastricht University Medical Center+, Maastricht, The Netherlands
Kempen, Laura ✱; Université de Liège - ULiège > Département des sciences fonctionnelles (DSF)
Lu, Chang; Cardiovascular Research Institute Maastricht, Experimental Vascular Pathology, Department of Pathology, Maastricht University Medical Center+, Maastricht, The Netherlands, Institute for Computational Biomedicine, Heidelberg, Germany
Donners, Marjo M P C; Cardiovascular Research Institute Maastricht, Experimental Vascular Pathology, Department of Pathology, Maastricht University Medical Center+, Maastricht, The Netherlands
Biessen, Erik A L; 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
Goossens, Pieter ; Cardiovascular Research Institute Maastricht, Experimental Vascular Pathology, Department of Pathology, Maastricht University Medical Center+, Maastricht, The Netherlands. Electronic address: pieter.goossens@maastrichtuniversity.nl
✱ These authors have contributed equally to this work.
Language :
English
Title :
Protocol for multispectral imaging on cryosections to map myeloid cell heterogeneity in its spatial context.
Hartstichting [NL] CSC - China Scholarship Council [CN]
Funding text :
This work has been supported by the European Research Area Network Joint Transnational Call for Cardiovascular Disease (ERA-CVD), the Dutch Heart Foundation ( JTC-2017t100 AtheroMacHete to P.G. and E.A.L.B. and Dekker 2020T042 to P.G.), and the China Scholarship Council (CSC) (no. 201706990018 to C.L.).
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