Shared Resource: Flow Cytometry

December 16, 2014
By: Wiiliam Alexander, Writer, DCI

Geoff Markowitz, a PhD candidate in cancer biology, puts to use a self-use analyzer, one of many available to researchers through the Flow Cytometry Shared Resource. Markowitz is studying liver cancer and is characterizing cells to better understand its progression in mouse models.Thirty-five years after its founding, the Duke Cancer Institute’s oldest shared resource remains as important as ever. Established in 1979, the DCI’s Flow Cytometry Shared Resource (FCSR) offers cell sorting and flow cytometric analysis for members of the DCI as well as the broader health system.

Located in rooms 306, 307, 336, 343, and 369 of the Edwin Jones Cancer Research Building, and rooms C313 and C316 of the Levine Sciences Research Center, the FCSR operates, maintains and upgrades instrumentation for flow cytometric analysis and cell sorting of cells prepared by investigators and brought to the FCSR. The FCSR provides four analyzers, four cell sorters and one image cytometer: these cytometers are available at all times to all DCI members.

As a 26-year veteran of the resource, FCSR co-director Michael Cook, PhD, takes part in all its daily activities, from performing cytometry to making repairs to preparing core grant renewals. Cook estimates the FCSR currently serves 300 to 400 Duke investigators a year, including faculty, fellows, medical and graduate students, and post-doctoral researchers.

The Flow Cytometry Resource team is headed up by Michael Cook, PhD, and Michael S. Krangel, PhD. Pictured from left to right: Nancy Martin, lab research analyst II; Michael Cook, PhD; and Bin Li, PhD, lab research analysts II. Not pictured: Lynn Martinek, lab research analyst II.“We have to know a little bit about the science, a little bit on how our computers and lasers work, and a little bit about human nature,” Cook said. “When I started here, we had two instruments and two employees,” he said.

“Now we have nine instruments and four employees, as well as excellent business support from the Department of Immunology. We have more employees, more instruments to perform the work, and the popularity of flow cytometry has increased exponentially in recent years,” Cook said.

The FCSR’s cytometers are capable of both analyzing and sorting cells–often at speeds of 10,000 cells per second or greater, Cook said. Cells that contain a component of particular interest for a researcher–such as a specific protein, DNA sequence or RNA molecule, can be measured, sorted and isolated.

Two of the cell sorters are four-laser instruments, one has three lasers, and one is a two-laser sorter. The staff provides state of the art multiparameter cell sorting of up to 12 simultaneous fluorochromes, up to six simultaneous populations, and level P2 biohazard containment. Researchers usually run the analyzers themselves or can have FCSR staff do run them. Fees are charged to partially offset operating expenses.

Qi-Jing Li’s, PhD laboratory uses the FCSR on a weekly, or even daily, basis.Qi-Jing Li, PhD, is one of the FCSR’s frequent patrons. His laboratory studies the role lymphocyte T-cells, part of the body’s immune system, play in recognizing and killing tumor cells. Finding these cells, however, is a challenge–less than 0.1 percent of the body’s T-cells have the capability to recognize and respond to tumors.

The FCSR’s cytometers have allowed Li’s laboratory to identify the receptors that recognize tumors and then engineer “whole armies” of T-cells that can recognize and fight tumors, Li said. His research now focuses on using these engineered T-cells to better locate and destroy tumors. As part of this work, Li visits the FCSR on a weekly, or even daily, basis.

“The FCSR staff are all very helpful–I know them all very well and they always find time for us,” Li said. “Mike never says no to helping us out.”

Read more about the FCSR at the FCSR’s website. To access the Flow Cytometry Shared Resource Facility registration form, click here. Once registered, most scheduling can be done online using a net ID and net ID password at cancerflowcore.duhs.duke.edu.

Circle photo: Nancy Martin, lab research analyst II, shares a laugh with Flow Cytometry Shared Resource user Thomas O’Brien, a postdoctoral associate in the Department of Immunology. Using the newest cell sorter, Martin purifies cell populations in preparation for cell sorting. Martin has been a member of the Flow Cytometry team for 13 years.