RE: [Histonet] storing frozen sections on glass slides in the freezer

From:"Hofecker, Jennifer L"

Hi Jason,
We do cut and store our frozen muscle slides in the freezer.  Our protocol is a little different that yours, but I will share anyway...
We cut our frozen sections of muscle at 8 microns and place on positive charged slides.  Sections are circled with a hydrophobic pen and placed in a slide box (one hundred slide plastic box).  The box is then placed in the  
-70 freezer.  We cut a few extra sections beyond what we need for our histochemistry panel and also store those in a slide box in the freezer.    We have gone back and pulled extra slides up to a few months later, and the enzyme histochems worked fine.  We also keep our normal muscle controls in a slide box at -70 and we pull them out as needed.  Again, no problems encountered with the staining.  As you pointed out in your post, be sure to allow slides to come to room temp before staining (we learned that the hard way).  
I realize that we're talking different tissue and different slides, but I wanted to let you know that we do this very successfully.  The main difference in our protocols that I would worry about is the -20 freezer.  Does it cycle through defrost everynight?  If it does, you could still use it, just pack any extra space around the slides with cold packs.  We've done that from time to time when we couldn't get things into the -70  the cold packs keep the temps fairly stable.  Not sure about the "chewing gum" method either.  Let us know how that goes...
Thanks and Good luck.
Jennifer Hofecker, HT (ASCP)
Vanderbilt University Medical Center
Division of Neuropathology
(615) 343-0083
(615) 343-7089 fax


From: on behalf of Jason Potas
Sent: Thu 8/10/2006 8:23 AM
Subject: [Histonet] storing frozen sections on glass slides in the freezer

Hi everyone/anyone,

I would like to store frozen rat spinal cord sections in a -20 degree C
freezer. Does anyone have experience with the following method, ie does it

1-cut sections frozen onto gelatinized glass slide.
2-air dry overnight at room temperature (to remove all moisture)
3-wrap multiple glass slides inside aluminium foil together like a pack of
chewing gum
4-store in the freezer at -20 deg C
5-when required, removed one (or required number) slide(s) from the pack,
but without defrosting other slides
6-allow to reach room temperature and commence histochemistry as normal

does it work without stuffing up the slides or the tissue sticking on the
back of other slides. has someone done this?


Jason Potas (PhD)
Laboratorio de Neurobiologia Celular e Molecular
Instituto de Biofisica Carlos Chagas Filho - IBCCF
Universidade Federal do Rio de Janeiro
CCS bloco J sala J1-029 Ilha do Fundao
21941-590 Rio de Janeiro, RJ - Brasil
Tel: +5521 2280 4694
      +5521 2562 6554
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