In regard to Claude's message of Tuesday, Feb 18th >....Has anyone tried a protease/enzyme solution followed up by dilute hydrogen >peroxide to sterilize?.....comments, plus or minus. ... Most cell types we run on the FACScan or FACS Vantage are easily handled with 0.5% Household bleach (1:10 dilution from the Chlorox bottle). Immediately after the last sample, we make a short 30 second flush with water with high flow rate followed by bleach for 10 minutes. This is followed by 10 minutes of sterile water, 2 minutes with automatic dishwasher rinsing agent (Jet Dry in the USA, SUN Rinse Gloss by Lever in Europe) and a final 3 minutes of sterile water. This method is also used when cells are stained with AO, EB or PI. When we are running cells known to be sticky due to producing mucins, e.g., Intestinal Epithelial cells, we do a fast flush with water followed by Hyaluronidase enzyme for 10 minutes, then water, jet dry, etc as above. The Hyaluronidase is purchased from Sigma (Cat# H2126) and 8 ug are dissolved in 25 ml dH20. I always sonnicate my nozzle tips in the enzyme after IEL cells are sorted as a preventive measure. As for sterilizing the sorter, we routinely use 70% ETOH unless we were doing work with bacteria previously, in which case 0.5% bleach or 3-6% hydrogen peroxide or 20% Acetic Acid can be used. I have not observed any corrosive effects on metal parts or connectors in either instrument in 4 years of operation. Most metal parts are stainless steel or anodized protecting them from harsh chemicals. Steven Merlin University of Bern FACS Core Facility Murtenstraase 31 CH-3010 Bern, Switzerland Lab: ++41-31-632-8876 Office: ++41-31-632-9960 FAX: ++41-31-381-8764 e-mail: merlin@patho.unibe.ch
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