How to deal with ARIA contamination?

Rosson, Dan P drosson at utmem.edu
Wed Jan 30 14:17:51 EST 2008


Join the club! 
 
We developed contamination problems after about 6 months of service in
our Aria. I never had it in a MoFlo or Elite. The "prepare for asceptic
sort" procedure that BD gives you involves removing the filters and then
sterilizing the unit with 70% EtOH. The filters that they give you are
supplied sterile to BD by Pall and Whatman. However, BD puts non-sterile
connections on them and supplies them to you in bubble wrap. If you
autoclave the unit, the water-tight connections are destroyed. I tried
buzzing them in a Cesium irradiator for 10 times longer than the charts
said was necessary, but the damned bugs survived. I never figured out
why. BD says not to use EtOH on the filters especially the bubble
filter, but I chased down its manufacturer and it turns out to be a
Whatman AS 36 which is made of nylon and glass fiber. ( It doesn't take
a PhD in chemistry --which I am-- to tell you EtOH won't hurt it). The
Pall filter is also EtOH resistant (Pall doesn't make a filter that
isn't reisistant to EtOH). So a few weeks back I performed the "prepare
for asceptic sort" and didn't take the filters off. The instrument
didn't dissolve and things were sterile for a while after that.
Recently, however, a couple of tests came back contaminated and the
insidious part of it is it seems to take 4 or 5 days for the
contamination to grow up. Anyway, I'm re doing the sterilization
procedure as I write this.
 
Well my two cents worth in this is  to go ahead and run 70% EtOH through
the entire unit including filters at what ever frequency you think is
appropriate ( I'm thinking every week or two) and do frequent sterility
tests and thus deal with the problem that way. Additionally, I have
users that routinely use gentamycin in their cultures and they never get
hit with contamination. Of course that's just my contamination: yours
may be different.
 
 
Dan Rosson PhD
University of Tennessee

________________________________

From: akos.szilvasi at novartis.com [mailto:akos.szilvasi at novartis.com] 
Sent: Monday, January 28, 2008 1:56 PM
To: cyto-inbox
Subject: How to deal with ARIA contamination?



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 <mailto:akos.szilvasi at novartis.com> 

 



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