How to deal with ARIA contamination?

Simon Monard smonard at ibmc.up.pt
Tue Jan 29 19:46:00 EST 2008


Hi Akos

I had some probelms with my Aria for a while. Drove me nuts. I ended up doing a
few things. I ditched that bubble filter entirely. I filled all the tanks one
at a time with 10% bleach and did a ton of tank primes, basically trying to
give every part of the fluidics a good bleaching. I ran bleach though the
sheath path with the stream on for a while. Then rinced out the tanks real good
with DI water and did the same with DI water, running it though all the lines
for a while. Then I replaced all the fluid filters with new ones  (you could
rinse then really well and autoclave them, the connections can get a bit leaky
after autoclaving). I then ran though a bunch of autoclaved sheath fluid. I now
autoclave my sheathfluid and haven't had any contaminations since. Doing this
took most of the day. I had tried using less drastic measures before but the
contamination kept coming back. There are so many places bacteria can lurk in
the Aria there being so much tubing. The bubble filter needs to go as it never
gets sterilized if you isolate it when running ethanol as BD suggest. For many
years I had no contaminations at all with Vantages etc.
You do need to make sure you get rid of any traces of bleach as cells don't get
on very well with it. I make up my own sheath fluid as 10x stock, dilute it to
1x and autoclave it in the spare sheath tank.

Before asceptic sorts I run 70% ethanol though the sheath path for a while (not
the sheath filter) and do an bunch of fills after switching back to sheath
fluid. I have done many sorts now with no problems even without having
antibiotics in media afterwards.

Best of luck

Simon

Quoting
akos.szilvasi at novartis.com:

> Dear FLOWers,
>
> We have a less than two years old (Special Order - UV equipped) ARIA
> sorter that had an unrelenting sheath fluid contamination problem. The
> recurring mysterious contamination existed from the arrival of the sorter.
> We reported it multiple times. BD replaced a few parts of the fluidics
> system but it kept the bugs out of the sheath only for 4-6 weeks at the
> time. Filter replacements, bleaching the system works only temporarily.
> Our other Aria sorter, an older one, never had any such symptoms. We use
> them in an identical fashion.
>
> One lab suggested to follow their solution (this is not an isolated
> occurrence - not matter what they say) to fill the distilled water tank
> with 70% ethanol so that the fluidics shut down procedure would fill the
> lines with alcohol. That sounds good but no one could tell if this trick
> has some detrimental effect on any parts of the fluidics system. Have you
> used alcohol over night in the sorter? Did it cause any problem on the
> long run?
>
> Any other solution? BD offers to replace the whole fluidics system for $
> 13,000.
>
> Best regards,
> Akos
>
> __________________________
> Akos Szilvasi
> NIBRI Core Laboratory Services
> manager
> USCA, 601-5301
> Novartis Institutes for BioMedical
> Research, Inc.
> 100 Technology Square
> Cambridge, MA 02139
> USA
> Phone: +1 617 8717177
> Email : akos.szilvasi at novartis.com
>
>
>
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-- 
Simon Monard
Cytometry Lab Manager
Rua do Campo Alegre, 823,
4150-180 Porto - Portugal
Tel +351 226 074 900 ext 3102 . Fax +351 226 099 157
Cellphone # 919 036 680

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