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

Elias B. Wieland; Laura J.A.P. Kempen; Chang Lu; Marjo M.P.C. Donners; Erik A.L. Biessen; Pieter Goossens

doi:10.1016/j.xpro.2023.102601

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

Elias B. Wieland, Laura J.A.P. Kempen, Chang Lu, Marjo M.P.C. Donners, Erik A.L. Biessen, Pieter Goossens^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

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

Original language	English
Article number	102601
Number of pages	17
Journal	STAR protocols
Volume	4
Issue number	4
Early online date	23 Sept 2023
DOIs	https://doi.org/10.1016/j.xpro.2023.102601
Publication status	Published - 15 Dec 2023

Keywords

Antibody
Immunology
Microscopy
Molecular Biology
Single Cell

Access to Document

10.1016/j.xpro.2023.102601Licence: CC BY

Cite this

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title = "Protocol for multispectral imaging on cryosections to map myeloid cell heterogeneity in its spatial context",

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",

keywords = "Antibody, Immunology, Microscopy, Molecular Biology, Single Cell",

author = "Wieland, {Elias B.} and Kempen, {Laura J.A.P.} and Chang Lu and Donners, {Marjo M.P.C.} and Biessen, {Erik A.L.} and Pieter Goossens",

note = "Funding Information: 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.). Publisher Copyright: {\textcopyright} 2023 The Author(s)",

year = "2023",

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doi = "10.1016/j.xpro.2023.102601",

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T1 - Protocol for multispectral imaging on cryosections to map myeloid cell heterogeneity in its spatial context

AU - Wieland, Elias B.

AU - Kempen, Laura J.A.P.

AU - Lu, Chang

AU - Donners, Marjo M.P.C.

AU - Biessen, Erik A.L.

AU - Goossens, Pieter

N1 - Funding Information: 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.). Publisher Copyright: © 2023 The Author(s)

PY - 2023/12/15

Y1 - 2023/12/15

N2 - 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

AB - 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

KW - Antibody

KW - Immunology

KW - Microscopy

KW - Molecular Biology

KW - Single Cell

U2 - 10.1016/j.xpro.2023.102601

DO - 10.1016/j.xpro.2023.102601

M3 - Article

SN - 2666-1667

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JO - STAR protocols

JF - STAR protocols

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