Re: staining old tissue
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From: | "R.Wadley" <s9803537@pop3.unsw.edu.au> (by way of histonet) |
To: | histonet@histosearch.com |
Reply-To: | |
Content-Type: | text/plain; charset="us-ascii" |
Dear Rebecca,
One of the great problems with precutting & storing tissue is that you
expose the tissue to oxidation (sp) & reduction reactions with the
surrounding environment. This is also true of blocks. However, by the
time you have trimmed the surface of a block you are into the unaffected
tissue. While there are many people who can report that they have stored
unstained slides forever without detrimental effect, it depends on the
staining. H&E no problem, many histochemical & some immuno staining - no
problem. BUT, for the really important stuff - big problems. In some
cases the chemical reactions that have taken place can be reversed, ie
antgen retrieval (if your lucky). I was taught to always store blocks, &
only store unstained slides for short periods (weeks to months depending on
what they were for). Refrigeration slows chemical reactions it doesn't
prevent them. If you must store unstained slides & must get good results
from them I can only suggest placing them immediately in a container filled
with nitrogen gas, temperature of storage - probably cold.
Long term storage of tissue in fixative creates similar problems.
Regards
Rob W.
At 07:48 12/07/1999 -0500, you wrote:
>Hi. I've got some mouse spinal cord tissue that has been sitting on slides
>in a freezer for two years. I need to stain it for oligodendricytes with a
>cc-1 antibody. The procedure that was prepared workes beautifully on
>tissue that has been UNCUT in a freezer for a long time, and was cut only
>recently to practice for this stain. However, when the precut old tissue
>(the important stuff) is used with the same procedure, the background
>staining is terribly high. Oh yeah, it's developed with DAB. The
>oligodendricytes are too light to be seen clearly against the background
>stain, making them uncountable. Our head histotech thinks its due to
>freezer burn, or something similar to it. Does anyone have any ideas as to
>how I can salvage the rest of this tissue?
>Thanks,
>Rebecca LoVerso
>Dept. of Neuroscience
>The Ohio State University
R. Wadley, B.App.Sc. M.L.S, Grad.Dip.Sc.MM
Laboratory Manager
Cellular Analysis Facility
School of Microbiology & Immunology
UNSW, New South Wales, Australia, 2052
Ph (BH) +61 (2) 9385 3517
Ph (AH) +61 (2) 9555 1239
Fax +61 (2) 9385 1591
E-mail r.wadley@unsw.edu.au
www http://www.micro.unsw.edu.au/caf.html
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