Hello Flow-ers,
I am writing to ask if anyone can give any advice, or practical
insights on sorting a preparation of purified B cells. I ran
some recently,(the cells were at about 15 million/ml) and
had quite a bit of difficulty keeping the cells flowing; they
didn't clog the stream so much, I just had to keep increasing
the differential pressure to get them to go through. Cleaning
out the sample line seemed to help a bit. When I had finished
the sort, I had sorted through about 8 million cells, while the
investigator said there were about 28 million cells in the tube
(although he wasn't sure at what point in the staining he had last
counted them). My questions are: is there a cell concentration
that works well for sorting B cells? Should they be more dilute
to prevent cells sticking together? Is there a special way to
wash, and treat the cells to optimize sorting/fluidic conditions?
Any advice would be appreciated. Thanks.
jyetz-aldape@genetics.com
Joanne Yetz-Aldape
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