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Tip #51. Inexpensive mesh filters for removing aggregates.
Tired of paying $$ for BD Falcon tubes with 35 micron cell strainer caps [~$500/case, 08-771-23]? Instead, use Sefar nylon mesh. Comes on a cardboard tube like wrapping paper. Cut off strips and use a paper cutter to make tidy 1-2 cm squares. Place filter onto eppendorf tube or facs tube, and pipette cell suspension onto filter. Lasts for several year. Autoclavable.
41 micron, #NC9802695 at Fisher, $~70/yard
Note: pipette a little slower with the mesh till you get the feel. Hasty pipetting makes samples run off the side of the filter rather than thru it. |
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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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