Re: FACSAria

Hey Flowers--

While the Aria is not the magical sorter that lets you read Socrates as 
it washes away all the worries inherent to high speed cell sorting, it 
is very sensitive functional sorting platform that performs admirably in 
our hands.  Like most first generation instrument it had its share of 
quirks (see our drawer full of fried 70um nozzles), but they are all 
readily surmountable.  One of the obvious differences compared with 
classic Jet-in-Air arrangements is the nozzles and we have had success 
with both the original CerMet (which we still use) and the standard 
nozzles in combination with the new stream charging arrangement.

One of the major factors contributing to erratic stream behavior is the 
nozzle, or more accurately, a dirty and/or plugged nozzle.  I recently 
invested in a nicely illuminated dissection scope so I can closely 
inspect the nozzles and it has been worth every cent.

Our usual approach has been to visually inspect the nozzle if we can't 
immediately get good streams and we have found several interesting things:

-biological gunge (for lack of a better term) can accrue inside the 
nozzle and it compromises stable stream formation.

-o-ring flashing (trim from the o-rings) has been found to very securely 
nest itself inside the nozzle (sometimes actually pulling halfway 
through the orifice) and all the sonication and bleach in the world will 
not get it out.

-bad o-rings have been found that will never make a good seal.	Whether 
this is due to our mishandling or they arrive that way has yet to be 
determined.

I have found that marginally steady hands and a tuberculin syringe will 
solve the first two problems as I can tease out most of these 
obstructions and the nozzle will perform fine.	So before you set fire 
to your $300K sorter, go invest a couple of hundred bucks in a scope (or 
locate one near your lab) and you might find sorting a little easier.  
If you have any more specific questions, feel free to contact me.

al





David Adams wrote:

>Overall, I've found my Aria to be the best sorter in my lab (1 Aria, 2
>DiVas, 1 SE).	However, I am very particular when choosing my nozzles. 
>In the old days, they used to send them 2-5 at a time, and I could count
>on getting one to two tips that sorted well and the rest that were
>basically useless.  The machine would eat the tips over the course of a
>few months, of course until I performed the Parks mod and then the BD
>upgrade.  
>
>I've been sent several ceramic tips, but none of them seem to perform
>well.	I am currently using three covetously horded 100um gold tips and
>getting excellent sort performance even at the 20 psi where I do most of
>my sorting.  
>
>David Adams
>University of Michigan 
>Flow Cytometry Core
>
>  
>
>>>>John Kearney <jfk@uab.edu> 08/23/05 9:47 AM >>>
>>>>	    
>>>>
>We have had a FACSAria in the lab for > 2yrs. Well actually two since
>the
>first one was replaced. We continue to be plagued by the inability to
>sort
>accurately and consistently  acceptably clean populations of mouse T
>and B
>cells on this machine, particularly if they are small subsets. Despite
>various fixes and technical advice this machine has been a big
>disappointment given the pre-release claims for this BD machine. I was
>wondering if other FACSAria users have had similar problems and if so
>were
>they surmountable? Thanks John
>John F Kearney
>Professor of Microbiology
>378 Tumor Institute
>University of Alabama at Birmingham
>Birmingham Al 35294
>jfk@uab.edu 
>http://www.uab.edu/luckielab/ 
>Ph 205 934 6557 
>FAX 205 934 1875 
>Courier 
>378 Tumor Institute
>1824 6th Ave South 
>Birmingham Al 35294
>
>
>
>
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