Office for Research and Graduate Studies (ORAGS)

Flow Cytometry Core Facility
Center for Vaccine Development

Directed by Marcelo B. Sztein, M.D., Professor of Pediatrics and Chief, Cellular Immunology and Flow Cytometry Section, CVD.

Mission Statement

The primary goal of the flow cytometry/cell sorting facility is to ensure that University of Maryland investigators whose research projects require the use of a flow cytometer have access to such instrumentation. The flow cytometry laboratory personnel have the expertise to assist in the preparation of experimental protocols, as well as to appropriately and efficiently operate the instrumentation.

Measurements/Services

The measurements/services listed below are currently available at the Flow Cytometry Core Facility, Center for Vaccine Development, University of Maryland.

1. Characterization of cell subpopulations, including lymphoid cells, bearing activation, adhesion and homing molecules by multicolor flow cytometry (up to 5 simultaneous fluorochromes + forward and side light scatter parameters). These include, for example, subsets defined by the expression of CD3, CD4, CD8, CD14, CD16, CD19, CD45R0, CD45RA, CD11a (LFA-1), CD103 (HML-1, αE/β7), CD29, CD28, CD62L (L-selectin), TCR α/β, TCR γ/σ, etc. Monoclonal antibodies labeled with FITC (Fluorescein Isothiocyanate), PE (Phycoerythrin), ECD (Energy Coupled Dye = PE-Texas-Red conjugate), PerCP (Peridinin Chlorophyll Protein) and APC (Allophycocyanin) are routinely used.

2. Measurement of intracellular levels of cytokines and other molecules in defined cell populations by multicolor flow cytometry

3. Determination of the expression of cytokine and chemokine receptors in defined cell populations by multicolor flow cytometry

4. Cell cycle analysis of proliferative responses as determined by propidium iodide or 7-AAD staining.

5. Measurements early events during cell activation as determined by measuring intracellular Ca++ fluxes and membrane potential

6. Determination of apoptosis in individual cells as measured by TUNEL staining, simultaneous staining with Annexin V and propidium iodide, and subG0/G1 peak analysis using DNA dyes such as Hoechst, propidium iodide or 7-AAD.

7. Measurement of expression of green fluorescence protein (GFP) in transiently or stably transfected eukaryotic and prokaryotic cells.

8. Isolation of lymphoid, tumor and other cells, as well as bacteria, by flow cytometric cell sorting based on expression of GFP and other markers defined by monoclonal antibody staining.

9. Measurement of cell proliferation by PCNA, BrdU and Ki67 staining.

Current & Previous Users of the Facility

1. Dr. Barry Handwerger
   Professor and Associate Chair for Research
   Department of Medicine
   Phone: 410-706-6414
   Fax: 410-706-0321 voice mail: 66474

2. Dr. Alan Cross
   Professor
   Department of Medicine
   Phone: 410-328-2565
   Fax: 410-328-6896

3. Dr. Jim Galen
   Assistant Professor
   Department of Medicine
   Phone: (410) 706-5328
   Fax: (410) 706-6205

4. Dr. Daniel Sussman
   Research Assistant Professor
   School of Pharmacy
   Phone: (410) 706-8497
   Fax: (410) 706-0346

5. Dr. Alessio Fasano
   Professor and Head, Division of Pediatric Gastroenterology and Nutrition
   Department of Pediatrics
   Phone: (410) 328-0812
   Fax: (410) 328-1072

6. Dr. Jim Manos
   Research Associate
   Marine Biotechnology Center
   Phone: (410) 234-8877
   Fax: (410) 234-8896

7. Dr. J. Glenn Morris, Jr.
   Professor and Chair
   Department of Epidemiology
   Phone: (410) 706-4580
   Fax: (410) 706-4581