[Histonet] storage of cryosections
Gayle Callis
gcallis <@t> montana.edu
Tue Dec 14 10:04:37 CST 2004
Birthe,
Initially, how you snap freeze is the first defense against freezing
artefacts. Frozen section stored with a dessicant is a good idea keep the
sections dry during storage. The storage container must stay closed to
equilibrate FS to room temperature. Water condensation is the enemy. Be
sure to air dry the sections before going into storage also. A black
plastic 25 slide capacity box works well. Also, opening a box directly
from freezer and taking a few sections out, then returning unused sections
to freezer must be avoided due FS freeze thaw. Put as many slides as you
need per storage container for a staining session.
Drierite is anhydrous calcium chloride and has a tendency to exfoliate fine
CaCl2dust all over everything including your sections which bothered
me. Try 10 to 18 mesh silica gel, (Fisher# S161-500). The little beads
have blue indicator so you know when dessicant is depleted. Put Silica gel
in a nylon processing bags (Thermo Electron or StatLab) or embedding bags
aka tea bags from Fisher, fold over top, and staple on fold the keep bags
closed.
Temperature is critical for storage - if you have -80C available, use that
instead of -20C, and NEVER store sections in a self defrosting freezer
found in a refrigerator, freeze dry cycle is very damaging. Hopefully you
can store FS at temperature lower than
-20C to preserve antigenicity over a long period of time. It may work for
short term storage and robust antigens.
At 02:23 PM 12/13/2004, you wrote:
>HI.
>Do I actually have to add Drierite absorbent into the boxes with the cryo
>sections in the -20C freezer to avoid freezing artefacts?
>Thanks, birthe
>
>
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Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)
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