Re: [Histonet] prions and frozen sections
Ditto~~but absolutely no frozens allowed.
LuAnn Anderson HT(ASCP)
University of Minnesota
At 03:20 PM 2/7/2006, Gayle Callis wrote:
>Our laboratory is BSL2 for prion work, murine and hamster animal
>models - brain, spinal cord and other tissues.
>A microtomy room is dedicated to prion work, and also contains a
>cryostat dedicated to prion work ONLY whether the tissus is fixed or
>not prior to cryomicrotomy. There is no way we can totally, 100%
>decontaminate the cryostat for prions, BSL2 or not, and I have too
>many other people, including students and technicians working on
>another cryostat and have to worry about prion
>decontamination. The researcher bought and maintains his own
>cryostat and hopefully it has a long life as no repairman would go
>inside it now.
>For paraffin work (also cryostat), workers take the necessary
>precautions, gloves, safety glasses, disposable lab coats, etc. when
>sectioning paraffin blocks and frozen sections. The microtome,
>waterbath are also dedicated to prion, nothing else is cut on these.
>Water from waterbath is dumped into a carboy of 6N NaOH (sitting in
>a big tub to collect any spills) until it reaches a volume where the
>NaOH becomes 2N simply by dilution, all is allowed to sit. We used
>to use bleach, that has gone by the wayside in favor of the
>NaOH. Carboy is then picked up by our biohazard safety laboratory
>on campus. Dry waste, towels, wipes, etc are collected for
>incineration i.e paraffin trimmings also. If we worked with sheep
>or CWD, then we would put sticky floor mats in front of doorway, and
>under microtome area. Trimmings should stay in the room.
>Sounds like a very uptight attitude for handling prions, but better
>to take stricter, proper precautions now and then have to change later.
>Some people collect alcohols into bleach but when we did this, we
>noticed a huge heat reaction - that does NOT happen with NaOH so we
>opted for the latter just not not have the heat problem.
>Research Histopathology Supervisor
>Veterinary Molecular Biology
>Montana State University - Bozeman
>PO Box 173610
>Bozeman MT 59717-3610
>406 994-4303 (FAX)
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