High-Dimensional Flow Cytometry Analysis of Regulatory Receptors on Human T Cells, NK Cells, and NKT Cells

Ryosuke Nakagawa; Jason Brayer; Nicole Restrepo; James J Mulé; Adam W Mailloux

doi:10.1007/978-1-0716-0849-4_14

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High-Dimensional Flow Cytometry Analysis of Regulatory Receptors on Human T Cells, NK Cells, and NKT Cells

Journal article

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High-Dimensional Flow Cytometry Analysis of Regulatory Receptors on Human T Cells, NK Cells, and NKT Cells

Ryosuke Nakagawa, Jason Brayer, Nicole Restrepo, James J Mulé and Adam W Mailloux

Methods in molecular biology (Clifton, N.J.), Vol.2194, pp.255-290

2021

DOI: 10.1007/978-1-0716-0849-4_14

PMCID: PMC7744271

PMID: 32926371

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https://www.ncbi.nlm.nih.gov/pmc/articles/7744271View

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Abstract

The field of flow cytometry has witnessed rapid technological advancements in the last few decades. While the founding principles of fluorescent detection on cells (or particles) within a uniform fluid stream remains largely unchanged, the availability more sensitive cytometers with the ability to multiplex more and more florescent signals has resulted in very complex high-order assays. This results in the co-use of fluorophores with increased levels of emission overlap and/or spillover spreading than in years past and thus requires careful and well thought out planning for flow cytometry assay development. As an example, we present the development of a large 18-color (20 parameter) flow cytometry assay designed to take an in depth analysis of effector lymphocyte phenotypes, with careful attention to assay controls and panel design.

Antibodies

Flow Cytometry - methods

Humans

Immune Checkpoint Proteins - metabolism

Immunophenotyping - methods

Killer Cells, Natural - cytology

Killer Cells, Natural - immunology

Natural Killer T-Cells - cytology

Natural Killer T-Cells - immunology

Staining and Labeling - methods

Details

Title: Subtitle: High-Dimensional Flow Cytometry Analysis of Regulatory Receptors on Human T Cells, NK Cells, and NKT Cells
Creators: Ryosuke Nakagawa - Moffitt Cancer Center
Jason Brayer - Moffitt Cancer Center
Nicole Restrepo - Moffitt Cancer Center
James J Mulé - Moffitt Cancer Center
Adam W Mailloux - Moffitt Cancer Center
Resource Type: Journal article
Publication Details: Methods in molecular biology (Clifton, N.J.), Vol.2194, pp.255-290
DOI: 10.1007/978-1-0716-0849-4_14
PMID: 32926371
PMCID: PMC7744271
NLM abbreviation: Methods Mol Biol
eISBN: 1071608495; 9781071608494
ISSN: 1064-3745
eISSN: 1940-6029
Grant note: P30 CA076292 / NCI NIH HHS R01 CA148995 / NCI NIH HHS P50 CA168536 / NCI NIH HHS
Language: English
Date published: 2021
Academic Unit: Microbiology and Immunology
Record Identifier: 9984297315502771

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