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Tip #14. Stability of paraformaldehyde (PFA)-fixed samples.
In the old days, stained and fixed samples could be stored up to a week at 4 degrees. This is because fluors like FITC and PE are quite stable. By contrast, tandem conjugates (Cy5PE, Cy7PE, PE-TR, etc.) are not stable after fixation, and rapidly decay. For optimal detection, analyze samples within 12 hours of fixation. |
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Lisa Borghesi
Assistant Professor & Director, Flow Cytometry Facility
Department of Immunology
On the naughty habits of FLOWers... |
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1. What’s the future of flow cytometry?
Simplicity. Accessiblity. Ten years ago, each instrument needed a full time
technician to align the laser and perform hardware compensation. Doing two sorts
a day was a major task. Now, a single skilled technician can perform multiple
sorts. In another ten years we'll have personal sorters in each lab.
2. So, in other words, you'll be out of the Director's position in ten years?
Rather, instruments that are currently part of the facility will move to the
bench top. In ten years, the new core instrumentation will combine flow
cytometry and confocal imaging so we will be able to visualize each cell in a
dot plot.
3. What is the biggest problem facing the facility?
Bad habits of users. I had no idea! And here I am talking about bad habits that
affect the quality of individual data, not habits that affect other people. We
are going to be addressing this through a series of tutorials in the coming
months.
4. What is an example of a bad habit?
Duplicating old experiments in order to re-use compensation controls. Would you use the standard curve from last week's ELISA on tomorrow's unknowns?
5. How do you and Dewayne coordinate your roles in the facility?
I’ve never seen someone as patient and unflappable as Dewayne. I think it’s
because he has three kids! We make a good team. Whereas I get easily agitated
when equipment goes down, Dewayne is unperturbed. Proof - number of times I have
called Dewayne when he has been on vacation: 6. Number of times he has called me
when I'm away: 0.
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