Dear all, just wanted to thank you all and those who have suggested ways of decontaminating my vantage Diva. Summaries are below (13 replies): 1. We have both Vantage and Aria sorters. The aria is more prone and harder to decontaminate than the Vantage. You need to remove the filter from the tank and throw it away. Connect the sheath live directly to the tank. Put a couple of liters of fresh 10% bleach made with sterile water into the tank, then run this through the entire system for 1 hr. Put a filled tube of the same bleach on the sample holder and put this in run. After the 1 hour turn off and leave it overnight.The next day. Rinse the tank with 70% ETOH several times to get the bleach out. Install a new filter (etoh will not harm filter). Then fill system and let run for 1 hour. Drain the filter of ETOH then rinse the tank with sterile DPBS. Fill tanks and start the system up. Run for 1 hour to get ETOH out of the lines. You are ready. Remember to use the bubble drain line to fill system between reagents. This is the best way to rapidly flush out the previous stuff. On a weekly basis, usually on Friday, the filter is drained into the tank and the tank is filled with ETOH. The fluidics is started up and there you go. We hate using bleach and only use it when we have positive cultures obtained from our systems. We have been known to fill the system with DPBS with 10 the antibiotics and let this set overnight. For rare persistent contaminations in our Aria. 2. 100% is not as lethal as 70% ethanol, yet NEITHER will kill everything. It's also most effective if you let it evaporate to dryness, which you can't do inside you sorter! You can use BD's built-in procedure which uses 10% bleach first (or FACS Clean), then the 70% EtOH to remove the bleach residue. Use autoclaved deionized water and sterile sheath ONLY in the other tanks as well. Use BD's Ethanol shut-down daily, too. Antibiotics will NOT kill those that are resistant, so it is NOT a good idea to EVER use them! You want a STERILE machine, but you want your cells of interest to survive the trip through the machine, too. --------------------------------------------- 3. One thing I would suggest is using 70% ethanol rather than 100%. For various reasons this has far higher bacteriocidal activity than 100%. An old trick I was shown is run baby bottle sterilising solution (eg Milton fluid) is addition. Its possible that you have a “slime” type culture in one of the nooks and crannies of the DiVa tubing, to be honest the only way to get rid of that is retube the machine although running a detergent like Coulter Clenz for an hour or two might help. In the short term you could try putting a .45micron syringe filter in the sheath line just before it reaches the head. If the problem is upstream of that you should clean things up a bit. 4. A couple of points, leaving 100% ethanol in overnight may make the tubbing brittle, 70% ethanol is supposed to be better at killing cells than 100%--_0a6cb2ca-80c7-4af0-94d8-957637416383_--Received on Wed Jun 4 14:58:00 2008
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