Re: Frozen problem
|From:||Aidan Schurr <Aidan.Schurr@hvh.co.nz>|
I orient my tissue on a large watch glass. By repeatedly picking up and placing the specimen on the glass, most of the liquid is left on the glass, without drying the specimen. It using takes 6 - 8 "placings" before sufficient liquid is removed. I have never had a problem since adopting this method (discovered by accident, as all the good ones are!!)
Hope this helps!
a i d a n c s c h u r r
section head, histology
department of pathology
hutt valley health
high street, lower hutt
telephone ++64 4 5709173
facsimile ++64 4 5709214
>>> Lynn Gardner <firstname.lastname@example.org> 12:05:00 p.m. Friday, 12 January 2001 >>>
Thanks Aidan, I thought about that earlier today and think this may be the
problem as we used PBS in place of saline and we used to use saline. How
are you drying or removing the transport media from the tissue without
drying out the tissue?
Thanks every so,
At 08:55 AM 1/12/01 +1300, you wrote:
>I have had a similar(ish) problem with my small skin biopsies for
>immunofluorescence. They come to me in transport media, and I would wash
>them in my DIF wash solution before freezing and embedding. The problem I
>discovered was that if there was significant amounts of the wash solution
>left on the outside of the specimen, this solution formed a "weak link" when
>sectioning, and it was impossible to get a section - all I would get was (as
>you say) a section of OCT with a hole in it. Do your muscle biopsies come
>to you in saline? If so, you may be experiencing a similar effect to me!
>Best of luck
>Aidan, New Zealand.
>a i d a n c s c h u r r
> section head, histology
> department of pathology
>hutt valley health
>high street, lower hutt
>telephone ++64 4 5709173
>facsimile ++64 4 5709214
>>>> Lynn Gardner <email@example.com> 4:43:46 a.m. Friday, 12 January
>We experienced an interesting problem when working with some muscle tissue
>the other day and was wondering if anyone out there has experienced it and
>if so how did they correct for it.
>We frozen some muscle tissue in isopentane cooled in liquid nitrogen to a
>temperature of -160. The tissue looked compeletly frozen when removed. We
>immediately placed the tissue in a vial and put in the -80 freezer. We
>embedded in OCT and tried to cut and all we got was a big hole in the OCT.
>The tissue seemed as though it was not frozen. Any idea of what happened??
>We have done this with other tissue and it worked fine. We know the freezer
>did not thaw out. So we are at a loss.
>I would appreciate any input! Thanks in advance gang!
>Lynn Gardner, HT(ASCP)
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