Office for Research and Graduate Studies (ORAGS)
Flow Cytometry-GCC
Scientific Objectives:
To provide state-of-the-art flow cytometry services for investigators in the program in oncology and campus wide at the University of Maryland School of Medicine.
Personnel:
Dean L. Mann M.D. Director
The cancer center flow cytometry facility was established as a shared resource in 1998 when Dr. Mann became director. He has more than 25 years experience in flow cytometry and was co-director of the first flow cytometry facility at the NCI, NIH. Over the years he has establish collaborations with numerous investigators at the NIH and more recently at the University of Maryland employing this technology to generate data that has been published in peer reviewed scientific articles.
Neil J Hardegen B.S. Program Director
Mr. Hardegen assumed his current position at the University of Maryland in January 2005. From 1993 to the end of 2004 he was manager of the Fluorescence Activated Cell Sorter and Imaging Core at the National Institute of Dental and Cranial- facial Research, NIH, Bethesda MD. There he was responsible for all aspects of flow cytometry including preparation of probes, data acquisition, multi-parameter cell sorting and data analysis. He was also responsible for training of professional and technical staff on the use of flow cytometric instrumentation and assisting with experimental design. Mr. Hardegen is trained on a variety of flow cytometry instruments made by different companies.
Dayton Nance B.S. Research Assistant
Mr. Nance was first employed as a Research Assistant in 1998-2000 conducting experiments that required flow cytometric expertise. From April 2002 to June 2003 he was employed full time in the Cancer Center flow cytometry core receiving formal training by BD in the use of their analytic and cell sorting instrumentation. He rejoined the Core operation as a part time employee in April 2004 and full time in August of that year. He has expertise in all aspects of multi-parameter flow cytometry data acquisition and cell sorting.
Location: Room 7-046, Bressler Research Building
Phone: 8-3915/16
Fax: 8-2822
Email: dnance@som.umaryland.edu or nhardegen@som.umaryland.edu
Web Site: http://www.umm.edu/cancer/research/flow_cytometry.html
Hours: Monday through Friday 7:30am – 4:30pm
Staff (areas of expertise):
Dean L. Mann (all services) dmann001@umaryland.edu
Neil J. Hardegen (all services) nhardegen@som.umaryland.edu
Dayton Nance (all services) dnance@som.umaryland.edu
Faculty Advisory Committee:
Douglas Ross, M.D., Ph.D., Professor, Medicine
Thomas MacVittie, Ph.D., Professor, Radiation Oncology
Peter Melera, Ph.D., Professor, Biochemistry and Molecular Biology
Miriam Smyth, Ph.D., Assistant Professor, Pathology
David Scott, Ph.D., Professor, Surgery
Dean Mann, M.D., Professor of Pathology and Director of Immunogenetics
Neil Hardegen, B.S., Program Director, Flow Cytometry Facility
Satisfied Customers: Laure Aurelian, Ph.D., Kurtis Bachman, Ph.D., Elizabeth Balcer-Kubiczek, Ph.D., Ricardo Feldman, Ph.D., Martin Flajnik, Ph.D., Amy Fulton, Ph.D., Ronald Gartenhaus, Ph.D., Don Gill, Ph.D., Dhan Kalvakolanu, Ph.D., Dean Mann, MD, Stephen Meltzer, MD., Aaron Rapoport, MD., Fey Rassool, Ph.D., John Sauk, DDS., Paul Shapiro, Ph.D., Naoko Takebe, MD., Stefanie Vogel, Ph.D., Richard Zhao, Ph.D.
Services Turnaround Time Instrument
Type: Acquisition/Analysis
Immunophenotyptic Assays daily FACScan or BD-LSR
Proliferation Assays daily FACScan or BD-LSR
DNA Cell Cycle daily FACScan or BD-LSR
Apoptosis Assays daily FACScan or BD-LSR
Stem Cell Discrimination daily Vantage or BD-LSR
Special Requests daily Depends on request
Type: Cell Sorting/Analysis
Sterile or nonsterile daily Vantage SE or DIVA
Based upon immunophenotyptic staining daily Vantage SE or DIVA
Based upon fluorescent proteins daily Vantage SE or DIVA
Stem Cell Discrimination daily Vantage SE or DIVA
Special Requests daily Depends on request
Pricing (Effective July 1, 2005) FY06
Service Operator Fee
Data Acquisition Facility Operator $100/hr
Investigator $50/hr
Data Analysis Facility Operator $40/hr
Investigator $20/hr
Cell Sorting Facility Operator Only $150/hr
Instrumentation:
Bench top Analyzers:
The Shared Services Facility has one FACScan and one BD-LSR cytometer for use by both technician assisted and user independent data acquisition/analysis. The FACScan has a fixed 488nm argon laser and three-color capabilities. The BD-LSR has three lasers (Argon 488nm, Helium-Neon 633nm, and Helium-Cadmium 325nm for ultraviolet excitation). The LSR has six fluorescence detectors, multiple pulse processing capabilities, and parameter ratio capabilities (i.e. Calcium flux applications).
Cell Sorters:
The Facility has two cell sorters: A FACSVantage SE with the Turbosort module and a CloneCyte attachment for single cell cloning. This sorter is equipped with two lasers; one Krypton-Argon Ion mixed gas enterprise laser that provides a 488nm and 355nm excitations, and one Heliun-Neon laser tuned to 633nm. The sorter has six detectors available for use in complex analysis and cell sorting. The second sorter is an SE with digital electronics (FACSDiVa option). It is equipped with three lasers: one Argon-Ion 488nm, one Krypton-Ion 405nm and one Helium-Neon 633nm. The instrument has eight detectors, Turbosort capabilities, and the CloneCyte attachment for single cell cloning. It is also equipped with an aerosol containment system for sorting of potentially biohazardous samples.
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