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Tip #99. Bad habit: Exploiting the "Duplicate without data" function on DIVA to re-use compensation controls from another experiment.
Accurate compensation relies on the brightness of controls and on the antibody lot. Different lots of antibodies have different degrees of fluorescence (due to distinct fluorochrome:protein ratios). Duplicate old experiments and sure, you'll get the same proportion of CD3, CD4, CD8 and CD19 cells, the big populations that are easy to resolve. What you won't get is accurate, reproducible resolution of rare populations - Tregs, tetramer positive cells, hematopoietic progenitors… |
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Dewayne Falkner
Manager, Flow Cytometry Facility
Department of Immunology
On the transition from bench to Facility Director... |
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1. How did you become facility manager?
When I heard that the Immunology Department was purchasing a flow cytometer I asked Penny Morel if I could be the one to go to training. I worked part time in the facility until there was enough work to keep me busy full time. With the help of Penny [whose grant writing efforts funded the Aria] and Olja [Department Chair] I transitioned from the lab to facility manager.
2. What is your vision for the flow cytometry facility?
I would like the facility to be a place where results are easily acquired and a resource for flow cytometric information.
3. What are the main challenges with running a core facility?
The facility has been growing since the LSR II was installed in 2003. I am still working on knowing what the users want and expect.
4. Do you miss life at the bench?
No!
5. When is the last time you loaded a sample on the flow cytometer?
What time is it now?
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