Nature Medicine
- 12, 972 - 977 (2006)
Published online: 23 July 2006; | doi:10.1038/nm1371
Quantum dot semiconductor nanocrystals for immunophenotyping by polychromatic flow cytometryPratip K Chattopadhyay1, David A Price1, Theresa F Harper2, Michael R Betts1, 3, Joanne Yu1, Emma Gostick4, Stephen P Perfetto1, Paul Goepfert5, Richard A Koup1, Stephen C De Rosa6, Marcel P Bruchez7 & Mario Roederer11
Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 40 Convent Drive, Bethesda, Maryland 20892, USA. 2
Solus Biosystems, 2454 Embarcadero Way, Palo Alto, California 94303, USA. 3
Department of Microbiology, University of Pennsylvania, 522E Johnson Pavilion, 3610 Hamilton Walk, Philadelphia, Pennsylvania 19104, USA. 4
Nuffield Department of Clinical Medicine, John Radcliffe Hospital, Oxford OX3 9DU, UK. 5
Department of Medicine, University of Alabama at Birmingham, 908 20th St. South, Birmingham, Alabama 35294, USA. 6
Fred Hutchinson Cancer Research Center, University of Washington, 1100 Fairview Avenue North, Seattle, Washington 98109, USA. 7
Department of Chemistry and Technology, Center for Networks and Pathways, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, Pennsylvania 15213, USA.
Correspondence should be addressed to Mario Roederer Roederer@nih.gov 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.
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