Date: Thu Jan 31 2008 - 09:45:49 EST
Hi Akos, I had the same problems. I use nitrogen gas from a 175 l LN2 Container for the auxiliary air supply. (The pressure of the LN2 contaiiner is set to 100 PSI) It is dry sterile and no O2 depending organisms will grow in the pressurized part of the instrument. I never used the bubble filter. During the long clean the plenums aren't completely filed with beach or 70% ETOH , so to get rid of the contamination I disconnected the Plenum tubing and filed the plenums completely with FACS Clean let them soak for a while and did the same with 70% ETOH and DI. Since I use HBSS without any preservative for sheet fluid I routinely run 70% Ethanol through the sheath path including the sheet filter (I replace the sheet filter every 6 months.) with the stream and sample line back flush on for a while twice and do the same with DI water and sheath fluid. It takes 60 minutes to start up the Aria but I don't have contaminations any more. Reinier van der Linden Erasmus University MGC- dept. of Cell Biology and Genetics (H Ee 1002) Dr. Molewaterplein 50 3015 GE Rotterdam the Netherlands Tel: +31-10-4087854 / +31-10-4087490 email: r.vanderlinden@erasmusmc.nl _____ From: akos.szilvasi@novartis.com [mailto:akos.szilvasi@novartis.com] Sent: Wednesday, January 30, 2008 15:09 To: cyto-inbox Subject: Re: How to deal with ARIA contamination? We did all of those you suggested, Simon. The air bubble filter landed in the garbage a long-long time ago. That dramatic bleaching (10% bleach in both the sheath and the dist. water tank followed by repeated start ups and shut downs) solved the contamination only temporarily, say, 4 weeks or so. Even in between we occasionally could grow a couple of colonies from the sheath sample. According to BD the house air can be a source of contamination. Strangely there is no barrier built in the outside compressed air source but (if I understood correctly) there is a filter for the sorter's own compressor. Next I will have to find an air filter for the house air (even though the line has two huge cases attached to - a moisture and an oil trap). Thanks for all the responses (on and off line). 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 : <mailto:akos.szilvasi@novartis.com> akos.szilvasi@novartis.com Simon Monard <smonard@ibmc.up.pt> 01/29/2008 08:08 PM To akos.szilvasi@novartis.com cc Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu> Subject Re: How to deal with ARIA contamination? 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@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@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. Thank you. > -- 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 ---------------------------------------------------------------- This message was sent using IMP, the Internet Messaging Program. _________________________ 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. 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