[Histonet] Mouse Tissue Problem

[Geoff McAuliffe]

Hi Judi

    All of the possible sources you mentioned could be the cause of your 
problems. I suggest re-embedding with fresh solutions for all steps. 
That will circumvent all of the problems exception fixation, which 
cannont be solved with this batch of tissue. In my experience, properly 
fixed and processed tissue should not have to be soaked for more than a 
minute or two to get good sections. However, mouse tissues processed on 
a processer with a "human tissue schedule" are often overly processed 
and wind up dry and brittle. I don't know what Telly's fixative is, 
perhaps a formalin-alcohol-acetic mix? Bouin's fixed material should cut 
beautifully.

Geoff

judi.ford <@t> jax.org wrote:

>Hi Everyone,
>
>Our lab has encountered a problem with a batch of mouse tissue processed recently. We receive tissue from various research labs and it is usually either fixed in Telly's or Bouin's for 24 hrs before processing. Nothing unusual was noticed until facing this batch of blocks. Some blocks faced fine while others, like muscle and tumors, shredded and the tissue was soft and spongy (light colored in the middle).  The blocks I have faced fine and were hydrated on an ice tray prior to sectioning.  In the past I've hydrated bouin's fixed blocks for a couple of days and never had a problem with cutting.  These blocks were hydrated in the frig overnight, which is my usual pattern, and then cut the next day.  I'm having a horrible time cutting the tissue. Intestines (not impacted with feces) are not cutting well (very compressed with a new knife) and the tissue breaks away from the paraffin both in the block and on the waterbath. The surface of the faced tissue appears wavy after hydration. My water bath temperature is 40 degrees centigrade.
>
>We've been discussing the following as possible causes: inadequate fixation, fixative mixed wrong, inadequate levels of solutions in the processor (although tissue on the top layer came out fine), tissue not cleared properly, etc. 
>
>I hoping someone may have some ideas on how to narrow down our troubleshooting with this problem.
>
>Thanks in advance.
>
>Judi Ford
>HIstotechnologist
>The Jackson Laboratory
>Bar Harbor, Me  04609
>
>
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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
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mcauliff <@t> umdnj.edu
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