Date: Fri Feb 01 2008 - 09:54:12 EST
I firmly beleive Mike is right! I came to UT about a year ago which was 6 months after they got the instrument. The autoclave cycle for the sheath fluid wasn't quite enough. Anyway, the first 12 months there were no contamination problems, then suddenly bugs all over the place! The filter had eventually developed pathways big enough for the bugs to get through. The rest of the story, I believe, is the post by Wayne Green which alludes to a study that concluded that there are pockets of sheath fluid that don't get flushed during a cleaning, thus it keeps coming back. The moral of the story seems to be don't let your instrument get contaminated in the first place. Alice Givans writes her instrument never had any contamination. I wonder if she would be willing to deliberately contaminate it to see if she could get it decontaminated again. ----Just a thought! Dan Rosson PhD University of Tennessee ________________________________ From: Michael Hughes [mailto:MHughes@picr.man.ac.uk] Sent: Friday, February 01, 2008 5:18 AM To: cyto-inbox Subject: RE: How to deal with ARIA contamination? Hi Dan, We have been running and Aria for about 3 years. Because we do sterile sorts every day we really needed to close all the doors to bacteria. The main problem we found was that the Pall filter on the sheath line eventually broke down, some quicker than others and so we replace it with a plastic millipore filter with a 0.22 micron filter we could replace every day with a freshly autoclaved one. We also replaced the bubble filter with a stainless steel Millipore filter which again we replace every day. We use this as backup just in case a filter splits. Regards, Mike Hughes, PICR, Manchester, England ________________________________ From: Rosson, Dan P [mailto:drosson@utmem.edu] Sent: Wednesday, January 30, 2008 7:18 PM To: cyto-inbox Subject: RE: How to deal with ARIA contamination? 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@novartis.com [mailto:akos.szilvasi@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@novartis.com <mailto:akos.szilvasi@novartis.com> _________________________ CONFIDENTIALITY NOTICE The information contained in this e-mail message is intended only for the exclusive use of the individual or entity named above and may contain information that is privileged, confidential or exempt from disclosure under applicable law. If the reader of this message is not the intended recipient, or the employee or agent responsible for delivery of the message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by e-mail and delete the material from any computer. 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