title: A Simple, Versatile Antibody-Based Barcoding Method for Flow Cytometry.
abstract: Barcoding of biological samples is a commonly used strategy to mark or identify individuals within a complex mixture. However, cell barcoding has not yet found wide use in flow cytometry that would benefit greatly from the ability to analyze pooled experimental samples simultaneously. This is due, in part, to technical and practical limitations of current fluorescent dye-based methods. In this study, we describe a simple, versatile barcoding strategy that relies on combinations of a single Ab conjugated to different fluorochromes and thus in principle can be integrated into any flow cytometry application. To demonstrate the efficacy of the approach, we describe the results of a variety of experiments using live cells as well as fixed and permeabilized cells. The results of these studies show that Ab-based barcoding provides a simple, practical method for identifying cells from individual samples pooled for analysis by flow cytometry that has broad applications in immunological research.
authors.affiliation: Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892; and.
authors.forename: Billur
authors.identifier: 0000-0002-6808-3776
authors.initials: B
authors.lastname: Akkaya
authors.affiliation: Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, MD 20852.
authors.forename: Pietro
authors.identifier: 0000-0003-3486-4061
authors.initials: P
authors.lastname: Miozzo
authors.affiliation: Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892; and.
authors.forename: Amanda H
authors.identifier: 0000-0003-4277-8411
authors.initials: AH
authors.lastname: Holstein
authors.affiliation: Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892; and.
authors.forename: Ethan M
authors.initials: EM
authors.lastname: Shevach
authors.affiliation: Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, MD 20852.
authors.forename: Susan K
authors.identifier: 0000-0001-7261-3437
authors.initials: SK
authors.lastname: Pierce
authors.affiliation: Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, MD 20852 munir.akkaya@nih.gov.
authors.forename: Munir
authors.identifier: 0000-0002-9949-9424
authors.initials: M
authors.lastname: Akkaya
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000145:classification / Q000302:isolation & purification*; D001330:Electronic Data Processing / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D051379:Mice; D013194:Staining and Labeling
chemical_list: D000906:Antibodies; D005456:Fluorescent Dyes