[Histonet] prions and frozen sections

[LuAnn Anderson]

Ditto~~but absolutely no frozens allowed.

LuAnn Anderson HT(ASCP)
Neuropathology Lab
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.
>
>
>
>
>Gayle Callis
>Research Histopathology Supervisor
>Veterinary Molecular Biology
>Montana State University - Bozeman
>PO Box 173610
>Bozeman MT 59717-3610
>406 994-6367
>406 994-4303 (FAX)
>
>
>
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