Stephen Perfetto, Ph.D.Vaccine Research Center40 Convent Drive, Room 5507Bethesda, MD, 20892-3015Phone: 301-594-8659Fax: 301-480-2788Sperfetto@nih.gov
Director, Flow Cytometry Core Section
The VRC Flow Cytometry Core Facility (FCC) works closely with all of the Immunology Laboratories at the VRC, managing a range of instruments from advanced analyzers to the most sophisticated sorters. The FCC serves the research community as well as the clinical trials supported by the VRC. This facility has become an internationally recognized entity, a world leader in the development and application of multidimensional and multicolor flow cytometry. The FCC has collaborated on or directly developed the necessary technologies to perform 18-color flow cytometry, analysis, and sorting. This technological feat required the combined development of novel antibody-conjugated fluorochromes, including nanocrystal development, hardware (the integration of multiple lasers and detectors, with the associated high-precision optics), software tools (for complex multidimensional data analysis), and validation techniques (for instrumentation and reagents quality control).
The FCC currently manages eight instruments with capabilities of serving a wide range of flow cytometric clinical and research demands:
The FCC facility leads the development and advancement of cytometric technology at all levels: instrumentation, reagents, detection, and analysis.
Stephen P. Perfetto received a B.S. degree in medical technology from the West Virginia University in 1977 and completed his M.S. degree in 1981 from West Virginia University. He studied and worked in the clinical blood bank and clinical immunology laboratories until 1988, when he joined the EPICS Division of Coulter Corporation. In 1990, he was recruited to the Walter Reed Army Institute of Research (WRAIR) and was the manager of the core flow cytometry facility. While at WRAIR, he was involved in large HIV vaccine trials and developed functional assays to study the immune system of infected individuals.
In 2000, he joined the VRC as a staff scientist and manager of the flow core facility. This facility is the world leader in multicolor flow cytometry and continues to actively develop this technology on a number of different fronts: hardware development and reagent and analysis development; analysis and presentation of metadata (for example, summarizing our functional analysis in which they have broken down each single response into hundreds of categories defined by the expression patterns of individual cytokines or other functional measurements); particular aspects of immune function or viral dynamics within the context of the major research efforts.
Perfetto SP, Ambrozak DR, Nguyen R, Roederer M, Koup RA, Holmes KL. Standard practice for cell sorting in a BSL-3 facility. Methods Mol Biol. 2011;699:449-69.
Wu X, Zhou T, Zhu J, Zhang B, Georgiev I, Wang C, Chen X, Longo NS, Louder M, McKee K, O'Dell S, Perfetto S, Schmidt SD, Shi W, Wu L, Yang Y, Yang ZY, Yang Z, Zhang Z, Bonsignori M, Crump JA, Kapiga SH, Sam NE, Haynes BF, Simek M, Burton DR, Koff WC, Doria-Rose NA, Connors M; NISC Comparative Sequencing Program, Mullikin JC, Nabel GJ, Roederer M, Shapiro L, Kwong PD, Mascola JR. Focused evolution of HIV-1 neutralizing antibodies revealed by structures and deep sequencing. Science. 2011 Sep 16;333(6049):1593-602.
Zarkowsky D, Lamoreaux L, Chattopadhyay P, Koup RA, Perfetto SP, Roederer M. Heavy metal contaminants can eliminate quantum dot fluorescence. Cytometry A. 2011 Jan;79(1):84-9.
Chattopadhyay PK, Perfetto SP, Yu J, Roederer M. The use of quantum dot nanocrystals in multicolor flow cytometry. Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2010 Jul-Aug;2(4):334-48.
Perfetto SP, Chattopadhyay PK, Lamoreaux L, Nguyen R, Ambrozak D, Koup RA, Roederer M. Amine-reactive dyes for dead cell discrimination in fixed samples. Curr Protoc Cytom. 2010 Jul;Chapter 9:Unit 9.34.
Perfetto SP, Ambrozak D, Nguyen R, Chattopadhyay P, Roederer M. Quality assurance for polychromatic flow cytometry. Nat Protoc. 2006;1(3):1522-30.
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Last Updated February 01, 2013