Research Flow Cytometry & Cell Sorting
Director: John F. Whitesides, PhD

Shared Resource Summary:
The DHVI Research Flow Cytometry Facility, led by John Whitesides, serves the analytical and cell sorting needs of the DHVI as well as support for researchers throughout the Duke Community. The Flow Cytometry Facility offers state-of-the-art cytometric support to DHVI investigators in Basic, Developmental, and Clinical Research. The Core consists of a Non-BSL-3 Flow Cytometry Facility and a BSL-3 Flow Cytometry Facility, which assist investigators in developing assays for measuring human immune responses to vaccines and infectious agents providing critical information about cell surface structure, identification, internal components and cell physiology. The Flow Cytometry Facility offers several cytometric services including BSL-1 through 3 live cell sorting, phenotypic acquisition, FRET (Fluorescent Resonance Energy Transfer), DNA cell cycle analysis and intracellular marker analysis. The analytical and sorting abilities exceed 8 simultaneous parameters and enable researchers to define subpopulations based on cell surface morphology as well as size and complexity. The DHVI Research Cytometry Facility is partially supported by the NIH and conforms with guidelines established to ensure that the facility has accessibility to the Duke Community. Our facilities’s goals are to supply our researchers with critical information about disease models that lead to direct applications related to clinical trials and human diseases and infections. Our facility also aids in the discovery of key elements in stem cell research, cardiovascular disease, bone marrow transplantation and immune reconstitution.
Personnel/Contact Information:
Director: John F. Whitesides, PhD Office Phone: (919) 684-4895 Lab Phone: (919) 684-4130 Fax: (919) 684-3282 Email: jwhtsds@duke.edu	Manager: Patti McDermott Office Phone: (919) 684-4130 Lab Phone: (919) 684-4130 Fax: (919) 684-2161 Email: pmcderm@duke.edu
Location: 4113 MSRBII, DUMC Box 103020, Durham, NC 27710
1029 GHRB, DUMC Box 103020, Durham, NC 27710
Getting Started:
Please see the Common Tools sidebar to register to use facilities, schedule or request a job, and to retrieve data. A Hazard Assessment Form needs to be completed for each NEW project to determine the appropriate level of containment and the SOP needed to assure proper safety precautions are used when working with your samples. These forms must be emailed to the DHVI Flow Facility Director or Manager at least 48 hours prior to starting a NEW project for review and approval. Hazard Assessment Form A Protocol Form is needed with every experiment to give the Facility Operators a description of your experiment to properly setup the assigned instrument. This form must be submitted to DHVI.Flow@notes.duke.edu before coming to your scheduled time. DHVI Flow Cytometry Protocol Form Current approved base rates are listed below for flow facility services. Discounts may apply to these rates based on membership in affiliated centers or programs. Please contact the DHVI Shared Resources Business Office at (919) 684-3349 or via email. DHVI Research Flow Rates (3-09)
Facilities and Instruments:
BD FACS Vantage SE (D01) – MSRBII 4038 BD FACS Aria (A01) – GHRB 1071 BD FACS Canto (C01) – GHRB 1057 BD LSRII (L01) - SANDS 302 BD LSRII (L02) – MSRBII 4111
Services Provided:
Experimental design consulting, protocol development and data analysis Independent and Operator Assisted acquisition of phenotype samples Operater Assisted sorting of prepared flow samples at Biosafety Levels 1, 2 and 3. One-on-one training in flow sample acquisition on FACS Canto and LSRII instruments One-on-one training in flow data analysis using FlowJo Software
Protocols and Documents:
Safety SOP for Cell Analysis and Sorting at BSL-1 and BSL-2 Safety SOP for cell analysis and sorting at BSL-3
Affiliated Centers:
Duke Radiation Countermeasures Center Grant - AI-067798 (radccore.mc.duke.edu) Duke University Center for Translational Research Grant - AI-051445 Southeast Regional Center of Excellence for Biodefense and Emerging Infections (www.serceb.org) Duke Center for AIDS Research (www.cfar.duke.edu) Center for HIV-AIDS Vaccine Immunology (www.chavi.org)
Publications:
2006-2008 Gasper-Smith N, Crossman DM, Whitesides JF, Mensali N, Ottinger JS, Plonk SG, Moody MA, Ferrari G, Weinhold KJ, Miller SE, Reich CF 3rd, Qin L, Self SG, Shaw GM, Denny TN, Jones LE, Pisetsky DS, Haynes BF. Induction of plasma (TRAIL), TNFR-2, Fas ligand, and plasma microparticles after human immunodeficiency virus type 1 (HIV-1) transmission: implications for HIV-1 vaccine design. J Virol . 2008 Aug. 82(15):7700-10. Epub 2008 May 28. Abstract Henry SC, Daniell X, Indaram M, Whitesides JF, Sempowski GD, Howell D, Oliver T, Taylor GA. Impaired macrophage function underscores susceptibility to Salmonella in mice lacking Irgm1 (LRG-47). J Immunol. 2007 Nov 15. 179(10): 6963-72. Abstract Chen BJ, Deoliveira D, Cui X, Le NT, Son J, Whitesides JF, Chao NJ. Inability of memory T cells to induce graft-versus-host disease is a result of an abortive alloresponse. Blood. 2007 Apr 1. 109(7):3115-23. Abstract Volkheimer AD, Weinberg JB, Beasley BE, Whitesides JF, Gockerman JP, Moore JO, Kelsoe G, Goodman BK, Levesque MC. Progressive immunoglobulin gene mutations in chronic lymphocytic leukemia: evidence for antigen-driven intraclonal diversification. Blood. 2007 Feb 15. 109(4):1559-67. Abstract Fecci PE, Mitchell DA, Whitesides JF, Xie W, Friedman AH, Archer GE, Herndon JE 2nd, Bigner DD, Dranoff G, Sampson JH. Increased regulatory T-cell fraction amidst a diminished CD4 compartment explains cellular immune defects in patients with malignant glioma. Cancer Res. 2006 Mar 15. 66(6):3294-302. Abstract Pubmed Archive