Cell Viability Multiplexing: Quantification of Cellular Viability by Barcode Flow Cytometry and Computational Analysis

Valentina Giudice; Victoria Fonseca; Carmine Selleri; Massimo Gadina

doi:10.1007/978-1-0716-3052-5_7

Cell Viability Multiplexing: Quantification of Cellular Viability by Barcode Flow Cytometry and Computational Analysis

Methods Mol Biol. 2023:2644:99-121. doi: 10.1007/978-1-0716-3052-5_7.

Authors

Valentina Giudice^{1

2}, Victoria Fonseca³, Carmine Selleri⁴, Massimo Gadina⁵

Affiliations

¹ Department of Medicine, Surgery and Dentistry "Scuola Medica Salernitana", University of Salerno, Baronissi, Salerno, Italy. [email protected].
² Cell Biology Section, Hematology Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, USA. [email protected].
³ Translational Immunology Section, Office of Science Technology (OST), National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS), National Institutes of Health, Bethesda, MD, USA.
⁴ Cell Biology Section, Hematology Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, USA.
⁵ Translational Immunology Section, Office of Science Technology (OST), National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS), National Institutes of Health, Bethesda, MD, USA. [email protected].

PMID: 37142918
DOI: 10.1007/978-1-0716-3052-5_7

Abstract

Fluorescent cell barcoding (FCB) is a useful flow cytometric technique for high-throughput multiplexed analyses and can minimize technical variations after preliminary optimization and validation of protocols. To date, FCB is widely used for measurement of phosphorylation status of certain proteins, while it can be also employed for cellular viability assessment. In this chapter, we describe the protocol to perform FCB combined with viability assessment on lymphocytes and monocytes using manual and computational analysis. We also provide recommendations for FCB protocol optimization and validation for clinical sample analysis.

Keywords: Fixable dyes; Flow cytometry; Fluorescent cell barcoding; Multiplexing; Viability.

MeSH terms

Cell Survival
Flow Cytometry / methods
Fluorescent Dyes*
Lymphocytes*

Substances

Fluorescent Dyes