[Histonet] paraffin sectioning-dry tissue?

[James Watson]

We use a 5% glycerin in denatured alcohol for our 100% alcohol on our tissue processor for routine animal tissues, this reduces the over dehydration of the animal tissue and greatly reduces the time required to soak the blocks.  Warning, if processing fat or cell pellets do not use this, we switch the containers to straight 100% reagent alcohol for them.  For fat we have a longer processing schedule and for cell pellets we have a short processing cycle.

James Watson HT  ASCP
GNF  Genomics Institute of the Novartis Research Foundation
Scientific Technical Leader II, Histology
Tel    858-332-4647
Fax   858-812-1915
jwatson <@t> gnf.org

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Caroline Miller
Sent: Thursday, February 05, 2015 6:41 AM
To: Geoff
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] paraffin sectioning-dry tissue?

Yes, exactly what Mike and Geoff said. All mouse tissue, especially liver, can be really dry and needs a 'soak'. I have left them for an hour before now but don't leave it for longer than 4 hours though because it can start to swell and de-process!

You will still only get a few non-chattery sections so be gentle. Thinner sections also help too (3-4.5). Plus low um polishing after you trim

Good luck! It is weird at first but you will get used to it!

Caroline

Sent from my iPhone

> On Feb 5, 2015, at 6:29 AM, Geoff <mcauliff <@t> rwjms.rutgers.edu> wrote:
> 
> This is common with mouse and rat tissues, they get "over-dried" with a typical processing schedule.
> Soaking the face of the block with a kimwipe wet with ice water for 60 -120 seconds will enable you to cut 10 nice sections, maybe more.
> 
> Geoff
> 
>> On 2/5/2015 6:23 AM, Emily Brown wrote:
>> Hello all!
>> 
>> I just started sectioning mouse liver in paraffin and the tissue is 
>> very dry.  I know it's not supposed to have water due to the 
>> processing, but the weird thing is that one tech's solution is to put 
>> a wet kimwipe on the block for a while.
>> It seems to me that there is a larger processing issue if this is 
>> happening, am I correct? And why add water when you've already 
>> dehydrated it?
>> Unfortunately, we do not have the set up to embed them ourselves, so 
>> we have to send them to a histology lab.  They were sectioning for 
>> us, but they are backlogged, so my boss wants me to do it.  
>> Therefore, I can't tell you how they were processed, but I think 
>> usually the histology lab manages to get good sections.
>> Is putting a wet kimwipe (using distilled water) the best way to get 
>> rid of chatter that's only in the tissue? The surrounding paraffin 
>> sections excellent.
>> This may have been answered already, but a very quick google search 
>> didn't help.  My googlefu is probably erratic as it's still early.
>> 
>> Emily
>> 
>> 
>> "By bitching and bitching and bitching, they could exhaust the drama 
>> of their own horror stories. Grow bored. Only then could they accept 
>> a new story for their lives. Move forward."
>> 
>> -Chuck Palahniuk, "Haunted"
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> 
> --
> --
> **********************************************
> Geoff McAuliffe, Ph.D.
> Neuroscience and Cell Biology
> Robert Wood Johnson Medical School
> 675 Hoes Lane, Piscataway, NJ 08854
> voice: (732) 235-4583; fax: -4029
> mcauliff <@t> rwjms.rutgers.edu
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