Re: FACSVantage

From: ntvan (ntvan@audumla.mdacc.tmc.edu)
Date: Wed Jun 30 1999 - 16:46:05 EST


We have 2 Facsvantages bought about 6-7 years ago.  We sorted extensively for both sterile and
non sterile sorts.  In the follows are sort speed, purity and yied for sorting Polyscience
Green spheres mixed with red spheres of 2 ucron and 5 micron diameter, respectively.  Purity
and yield determined by hemocytometry under blue light for excitation and green band pass
filter for observing fluorescent spheres.  CV is about 2%.  One operator was involved.

Sort speed/sec        Purity        Yield
500                        >99%        >90%
1000                        99%        80-90%
1500                        98%        60%
2000                        95-98%       40-50%
2500                        90%            30-40%

As can be seen the purity didn't seem to be affected adversely with high speed sorting but
yield suffered very significantly.  The reason was abortion rate increased from 5-10% to
40-50%.

With cell lines sorting rate could not be maintained above 1000/sec without having cells
clogging the nozzle tip. We use a 70 micron diameter tip.  A 100 micron tip might be better
for clogging but drop formation rate was reduced.

Cells from peripheral blood or bone marrow could be sorted as fast as sphere if an in line
filer (25 micron mesh) was used with comparable yield.  We however rarely sort above
1500-2000/secs.

Hpe that these information is useful to you


Cliff McArthur wrote:

> Hi Everyone:
>
> Thanks in advance for this:
> I have an investigator who is considering a major purchase, as purchases of
> high-speed sorters usually are...
> The request is this, for FACSVantage users:
> I would love to hear from you what you are routinely getting for yields,
> recovery and purity (using whatever definitions you'd like) on the fastest
> FACSVantage sorts.  I have never used a FACSVantage, so I do not know whether
> or not the TurboSort or other components are normally part of a FACSVantage.
> I am looking for results off of the fastest of FACSVantage configurations
> (aside, of course, from whatever special tinkering individuals may have done).
>  I would also appreciate hearing from users about what speed - stream
> velocity, pressure, and total event rates - at which you are running.
>
> Big thanks,
> Cliff McArthur
> UC San Francisco
> 415-502-6860
>
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