[Histonet] Frozen tissue question

[Emily Sours]

>From the research point of view, I've heard of people not fixing tissue
before they section it. Since I work with embryonic tissue (which is mostly
water!), we always fix our tissue. It definitely is better to not fix tissue
when you want to stain with antibodies because anything you do to the sample
might affect the antigens.  However, since it's impossible to section
unfixed tissue, you have to fix it somehow.
Our lab once tried to section snap frozen unfixed chick embryo because
another lab (our Sauron, if you will) had said that's the best way to get
antibody staining.  We, however, could not get the tissue to section
properly.  How that other lab did it is beyond me.
Either way, there's never a clear yes or no with protocols, it'll always
change due to what you're sectioning.

Emily

A great book should leave you with many experiences, and slightly exhausted.
You should live several lives while reading it.
-William Styron



On Thu, Jun 23, 2011 at 1:47 PM, Nicole Tatum <nicole <@t> dlcjax.com> wrote:

> I run a Mohs lab that processes skin by frozen section. There is no need
> to use any fixative before hand. But, if you are looking for melanoma or
> melanocytes, freezing can cause artifact and make it difficult to read
> slides. Limit the amount of nitrogen you use to freeze the specimen. Some
> places use alcohol as a fixative after the slide is cut. It will for all
> purposes remove the water continent from the tissue which will preserve
> tissue. Hope this helps.
>
> Nicole Tatum HT ASCP
>
>
>
>
>
>  We have a new doctor in our lab who swears that all frozen tissue must
> > be fixed in formalin with a subsequent sucrose treatment before freezing
> > in OCT because not fixing it will cause the structures to be distorted
> > and you can't get good antibody attachment. In my previous experience,
> > we have done this with tissue that came from an animal that was perfused
> > with formalin before the tissue was removed. However, the majority of my
> > previous frozen specimens were flash frozen in OCT and fixed after
> > sectioning. It is also my understanding that fixation in formalin can
> > cause poor antibody detection because of cross-linking caused by the
> > formalin. I would like to hear some other opinions on this please. The
> > particular specimen we will be working with is skin.
> >
> > Thanks in advance,
> > Joel Reichensperger
> >
> > --
> > Joel Reichensperger
> > Researcher II
> > Southern Illinois University
> > Plastic Surgery Institute
> > jreichensperger <@t> siumed.edu
> > 217-545-7309 (Office)
> > 217-545-1824 (Fax)
> >
> >
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