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


Journal

Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969

Informations de publication

Date de publication:
2021
Historique:
entrez: 14 9 2020
pubmed: 15 9 2020
medline: 1 4 2021
Statut: ppublish

Résumé

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.

Identifiants

pubmed: 32926371
doi: 10.1007/978-1-0716-0849-4_14
pmc: PMC7744271
mid: NIHMS1650769
doi:

Substances chimiques

Antibodies 0
Immune Checkpoint Proteins 0

Types de publication

Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

255-290

Subventions

Organisme : NCI NIH HHS
ID : P30 CA076292
Pays : United States
Organisme : NCI NIH HHS
ID : P50 CA168536
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA148995
Pays : United States

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Auteurs

Ryosuke Nakagawa (R)

Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA.

Jason Brayer (J)

Department of Malignant Hematology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA.

Nicole Restrepo (N)

Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA.

James J Mulé (JJ)

Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA.
Department of Cutaneous Oncology, H. Lee Moffitt Cancer Center, Tampa, FL, USA.

Adam W Mailloux (AW)

Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. adam.mailloux@moffitt.org.

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Classifications MeSH