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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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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web