Hi folks! Thanks to everyone who responded to my query about the handling of HIV+ and other biohazardous samples by flow cytometry. Here is a summary of the responses: (1) The general consensus is that Universal Precautions should be taken when handling biohaardous material under any circumstances. Sample fixation with at least 1% paraformaldehyde for at least 4 hours is generally considered sufficient to inactivate virus when analysis is carried out on a closed-flow-system instrument such as the B-D FACScan or Coulter XL. Published guidelines include Current Protocols in Cytometry page 5.1.12 (suggested by William Nostrum), CAP and NCCLS guidelines (Suggested by Andrea Illingsworth of Dahl-Chase) and MMRW vol 41(RR-8), 1992, MMRW vol. 42 (RR-3), 1994 and MMRW vol. 46 (RR-2), 1997 (suggested by Janet Nicholson of the CDC). (2) For handling samples with viable virus on stream-in-air sorting systems, containment of aerosol is absolutely essential. Janice Giorgi has described protocols for monitoring aerosol containment using T4 phage escape from the sample stream in Flow Cytometry (Methods in Cell Biology vol. 42, 2nd edition, Academic Press, 1994, Chapter 21, p. 359). Ingrid Schmidt will also have a chapter describing these methods in the upcoming edition of J. Paul Robinsons's Handbook of Flow Cytometry Methods. I also found some other sources of information, including earlier MMWR reports on handling of HIV- and HBV-contaminated material, and studies on the inactivation of virus using formaldehyde. I can give the references to anyone interested. Thanks again for the feedback. Bill Telford Flow Cytometry Core Facility Hospital for Special Surgery Cornell University Medical College New York, NY 10021
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