Quantum dot semiconductor nanocrystals for immunophenotyping by polychromatic flow cytometry.
Immune responses arise from a wide variety of cells expressing unique combinations of multiple cell-surface proteins. Detailed characterization is hampered, however, by limitations in available probes and instrumentation. Here, we use the unique spectral properties of semiconductor nanocrystals (quantum dots) to extend the capabilities of polychromatic flow cytometry to resolve 17 fluorescence emissions. We show the need for this power by analyzing, in detail, the phenotype of multiple antigen-specific T-cell populations, revealing variations within complex phenotypic patterns that would otherwise remain obscure. For example, T cells specific for distinct epitopes from one pathogen, and even those specific for the same epitope, can have markedly different phenotypes. The technology we describe, encompassing the detection of eight quantum dots in conjunction with conventional fluorophores, should expand the horizons of flow cytometry, as well as our ability to characterize the intricacies of both adaptive and innate cellular immune responses.
Chattopadhyay
Pratip K
PK
Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 40 Convent Drive, Bethesda, Maryland 20892, USA.
Price
David A
DA
Harper
Theresa F
TF
Betts
Michael R
MR
Yu
Joanne
J
Gostick
Emma
E
Perfetto
Stephen P
SP
Goepfert
Paul
P
Koup
Richard A
RA
De Rosa
Stephen C
SC
Bruchez
Marcel P
MP
Roederer
Mario
M
D017252:CD3 Complex / Q000276:immunology; D016827:CD8 Antigens / Q000276:immunology; D000939:Epitopes; D005434:Flow Cytometry / Q000379:methods*; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D006684:HLA-DR Antigens / Q000276:immunology; D006801:Humans; D016130:Immunophenotyping / Q000379:methods*; D008285:Major Histocompatibility Complex; D036103:Nanotechnology; D045663:Quantum Dots; D012666:Semiconductors; D013601:T-Lymphocytes / Q000276:immunology
D017252:CD3 Complex; D016827:CD8 Antigens; D000939:Epitopes; D005456:Fluorescent Dyes; D006684:HLA-DR Antigens; D016650:Fluorescein-5-isothiocyanate