Facility instrumentation includes a FACSCalibur, a FACS Aria Special Order System(SOS) a 4-laser LSR II analytical cytometer and a 4 laser sorter.
The FACSCalibur was purchased in 2000 and is a two-laser (488nm and 615nm), four-color, six-parameter, bench-top analyzer used primarily for evaluating surface and intracellular antigen expression, apoptosis detection, bead-based cytokine detection and cell-cycle analysis. Users are able to run their own samples on this instrument after training with the facility director.
The flow cytometry shared resource has two FACS Aria II SOS cell sorting systems. One of these was purchased in 2010 by the Cancer Center and one is available through the CRI. Both Arias are fully-digital, four laser, ten color, twelve-parameter instruments. The Cancer Center Aria is configured with 405nm, 488nm, 561nm, 640 nm lasers and is available for sorting BSL1 and standard BSL2 samples. The CRI Aria is configured with 405nm, 488nm, 532nm, 633 nm lasers and is housed in a negative pressure room and operated under enhanced safety precautions required for BSL2+ samples. Both instruments are used for cell sorting experiments for live cell samples and collection of up to four populations simultaneously or clonal/ single cell sorting into a 96-well plate formats. Both systems are equipped with aerosol management systems and temperature control options.
The LSR II analytical cytometer has four lasers; 405, 488, 532, and 633 nm. This analytical cytometer is fully digital, with ten color detection and twelve parameters. It has a high throughput system with a 96 well plate loader. The LSR II’s primary function is to characterize complex populations of cells. The LSR II can collect data at a rate up to 35,000 events per second, although it is typically used at much lower speeds. Users are able to run their own samples on this instrument after training with the facility staff.
Calvin Williams, MD, PhD
Dr. Williams joined MCW in 2002 and served as the Director of the CRI Flow Cytometry Core since its inception in 2005. Dr. Williams obtained his MD and PhD degrees from the University of California-Irvine in 1991, and has been active in research focused on the mechanisms of immunologic tolerance. Dr. Williams’ recent work involves the characterization of regulatory T cell development and function in models of inflammatory autoimmune disease. He also serves as the Chief Scientific Officer for the CRI and as an Associate Dean for Research at MCW.
Tamara Nelson, MS, Research Associate
Research Associate returned to MCW in 2013. She earned her graduate degree from
the University of Wisconsin-Madison Biotechnology in 2007. With over ten years of research experience in both academic and biotech industry, she is well versed in many molecular biology and immunochemical techniques. Ms. Nelson provides training for new users, schedules and executes cell sorting, maintains and calibrates the instruments, and manages data archiving for the shared facility. She has completed coursework at BD Biosciences to help troubleshoot sorting rare cell types, small and large cells, as well as maximizing sort optimization and cell recovery.