[Histonet] Paraffin liver sections

[Victoria Baker]

Sorry - short addenda - depending on what they were doing with this tissue
may also be critical as over processing or extended fixation may affect the
tissue microscopic evaluation.

Vikki

On Fri, Nov 5, 2010 at 7:37 AM, Victoria Baker <bakevictoria <@t> gmail.com>wrote:

> Hi -
>
> Whole mouse livers do not need to be fixed for 72 hours given the process
> you described.  The protocol I followed for fixation of whole mouse organs
> was within 24 hours, also when processing I started at 70% etoh, I also
> included a 50/50 ratio of abs/xylene mix for my first clearing as it was not
> as harsh on smaller tissue samples.  The times and heat on a processor will
> turn this tissue very brittle if not handled correctly.  If you can get a
> better description of his processing this would be a help.
>
> When cutting trimming and soaking on ice will help, but if you put the
> block into warm water for about 30 to 45 seconds and then put it onto an ice
> slurry will also rehydrate the block enough to get a decent section as
> well.
>
> I hope this helps.
>
> Vikki
>
>   On Thu, Nov 4, 2010 at 2:08 PM, Adam . <anonwums1 <@t> gmail.com> wrote:
>
>> Dear all,
>>
>> One of my colleagues has consulted me about some paraffin embedded mouse
>> livers he's been trying to cut. They were fixed in neutral buffered
>> formalin
>> for 3 days and then processed and embedded into paraffin. He was trying to
>> cut 5 uM sections on our microtome, except as soon as the blade hit the
>> liver, the liver would start to roll up and collapse into a small sliver,
>> and you'd just get sections of paraffin with a hole where the liver should
>> be. We tried changing cutting angle and soaking the blocks in ice water
>> with
>> no success. This is the first time we've tried livers, but we routinely
>> cut
>> decalcified bones on that microtome and have never seen that. Any ideas on
>> what is going on?
>>
>> Thanks,
>> Adam
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>
>