Details

Autor(en) / Beteiligte

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

Titel

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

Ist Teil von

• STAR protocols, 2023-12, Vol.4 (4), p.102601-102601, Article 102601

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2023

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• 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 [Display omitted] •Single-shot approach for immunofluorescent multiplex imaging•A pipeline for spectral library composition before multispectral imaging•Various approaches to reduce tissue autofluorescence including bleaching•An open-access script to allow alignment of histological and immunofluorescent images Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.