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Dr. Mike Borowitz
Dr. William G. Telford
Bill Telford received his Ph.D. in microbiology from the Michigan State University in 1994, where his laboratory developed some of the earliest techniques for flow cytometric detection of apoptosis. He was appointed Staff Scientist at the National Cancer Institute, National Institutes of Health in 1999, and is currently an Associate Scientist and director of the flow cytometry core laboratory in the NCI Experimental Transplantation and Immunology Branch.
Lecture and Wet Lab
Flow cytometry plays a critical role in the identification and characterization of mammalian and human stem cells. We will review recent advances in stem cell analysis by flow cytometry, including both traditional and novel cell surface phenotyping approaches; physiological characteristics such as ABC transporter activity using pump-specific efflux probes and pump-specific antibodies; and flow cytometric measurement of other stem cell-specific physiological characteristics, including aldehyde dehydrogenase activity. Modern application to stem cells and recent technological innovations will be emphasized. We will do either a Hoechst 33342 or DCV side population labeling of mouse bone marrow, combined with cell surface labeling with three to six markers, allowing identification of SP activity in both early hematopoetic stem cells (HSCs) and more committed common lymphoid progenitors (CLPs). We will use the resulting polycolor samples to demonstrate the acquisition and analysis of complex multicolor samples, particular detector setup and both real-time and post-acquisition compensation setup using both the BD DiVa software and third-party packages like FlowJo.
Telford, W.G. (2010) Stem cell side population analysis and sorting using DyeCycle Violet. In Current Protocols in Cytometry, Robinson, J.P., Darzynkiewicz, Z., Dobrucki, J., Hoffman, R.A., Nolan, J.P., Orfao, A., Rabinovitch, P.S., eds., John Wiley and Sons, New York, NY. Chap. 9, Unit 9.30.
Telford, W.G., Bradford, J, Godfrey, W., Robey, R.W. and Bates, S.E. (2007) Side population analysis using a violet-excited cell permeable DNA binding dye. Stem Cells 25, 1029-1036.
Cabana, R., Frolova, E.G., Thomas, R.A., Krishan, A. and Telford, W.G. (2006). The minimal instrumentation requirements for Hoechst side population analysis: Stem cell analysis on low-cost flow cytometry platforms. Stem Cells 24:2573-2581.
Telford WG, Frolova EG. (2004) Discrimination of Hoechst side population in mouse bone marrow with violet and near-UV laser diodes. Cytometry 57A: 45-52.