Protocol for high-plex, whole-slide imaging of human formalin-fixed paraffin-embedded tissue using PhenoCycler-Fusion.

Autor:	Donovan ML; Queensland Spatial Biology Centre, Wesley Research Institute, Level 8 East Wing, The Wesley Hospital, Auchenflower, QLD 4066, Australia; Frazer Institute, Faculty of Medicine, The University of Queensland, Brisbane, QLD 4102, Australia., Jhaveri N; Akoya Biosciences, The Spatial Biology Company, Marlborough, MA, USA., Ma N; Akoya Biosciences, The Spatial Biology Company, Marlborough, MA, USA., Cheikh BB; Akoya Biosciences, The Spatial Biology Company, Marlborough, MA, USA., DeRosa J; Akoya Biosciences, The Spatial Biology Company, Marlborough, MA, USA., Mihani R; Akoya Biosciences, The Spatial Biology Company, Marlborough, MA, USA., Berrell N; Queensland Spatial Biology Centre, Wesley Research Institute, Level 8 East Wing, The Wesley Hospital, Auchenflower, QLD 4066, Australia; Frazer Institute, Faculty of Medicine, The University of Queensland, Brisbane, QLD 4102, Australia., Suen JY; Critical Care Research Group, The Prince Charles Hospital, Brisbane, QLD 4032, Australia; Institute for Molecular Bioscience, The University of Queensland, Brisbane, QLD 4072, Australia., Monkman J; Frazer Institute, Faculty of Medicine, The University of Queensland, Brisbane, QLD 4102, Australia., Fraser JF; Queensland Spatial Biology Centre, Wesley Research Institute, Level 8 East Wing, The Wesley Hospital, Auchenflower, QLD 4066, Australia; Critical Care Research Group, The Prince Charles Hospital, Brisbane, QLD 4032, Australia; Institute for Molecular Bioscience, The University of Queensland, Brisbane, QLD 4072, Australia., Kulasinghe A; Queensland Spatial Biology Centre, Wesley Research Institute, Level 8 East Wing, The Wesley Hospital, Auchenflower, QLD 4066, Australia; Frazer Institute, Faculty of Medicine, The University of Queensland, Brisbane, QLD 4102, Australia. Electronic address: arutha.kulasinghe@uq.edu.au.
Jazyk:	angličtina
Zdroj:	STAR protocols [STAR Protoc] 2024 Sep 20; Vol. 5 (3), pp. 103226. Date of Electronic Publication: 2024 Jul 18.
DOI:	10.1016/j.xpro.2024.103226
Abstrakt:	Single-cell spatial analysis of proteins is rapidly becoming increasingly important in revealing biological insights. Here, we present a protocol for automated high-plex multi-slide immunofluorescence staining and imaging of human head and neck cancer formalin-fixed paraffin-embedded (FFPE) sections using PhenoCycler-Fusion 2.0 technology. We describe steps for preparing human head and neck cancer FFPE tissues, staining with a panel of immunophenotyping markers, and Flow Cell assembly. We then detail procedures for setting up for a PhenoCycler-Fusion run, post-run Flow Cell removal, and downstream analyses. For complete details on the use and execution of this protocol, please refer to Jhaveri et al. 1 . Competing Interests: Declaration of interests N.J., N.M., B.B.C., J.D., and R.M. are employees of Akoya Biosciences, Inc. (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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