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I have taken a job change and now am responsible for all flow cytometry at another
company, and need some quick opinions and advice.
Where I once isolated cells, THEN stained them, I now hear that for PBLs anyway, the
recommended procedure is to stain with whole blood. I found the lab full of antibodies
from B-D and Immunotech that describe a procedure whereby 100 ul of blood are added to
20 ul of antibody...
Since I now, unlike before, work with SIV-infected blood and tissues, I have also been
told that it is recommended that with infected blood, [time-consuming] 12x75mm tubes are
used for the staining procedure as opposed 96 well microplates.
My questions:
1- Is it entirely true that it is best not to isolate the PBLs before staining? It would
be SO much nicer, and take much less time in the P3 lab if I could ISOLATE, FIX
overnight in the regular lab, then STAIN at my [careful]leisure in PLATES with VACUUM
and in normal clothes (what a wish-list!).
2- Can someone recommend reference material, on and off the 'Net, that I can use/buy?
3- I have a FACSort that has a Bering hard drive which is connected to bernoulli-150s on
a PC. The have had compatibility problems in the past and I have the go-ahead to update
this set-up. The first thing I see that is needed is to have a data backup separate from
the PC and leave the bernoullis with IT. I came from using a [antiquated as I am
hearing] FACSCAN so the FACsort appears to be stat-of-the-art to me ;-} Does anyone have
an opinion on this?
I know that these things have been written of in previous threads, so private e-mail
would be appreciated if any of you potential contributors believe it to be redundant,
but as I no longer have my old account containing the old messages, I cannot review
them!
Your experiences and advise is appreciated!
P. Echeagaray
Clinical Virology/Flow Cytometry
Frederick Research Center/Southern Research Institute
Frederick MD
301-694-3232 ext 111
plment@worldnet.att.net
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