[Histonet] Cryostat issues with lobster tissues

From:Norman Meres



Hello:
	I am adapting a histochemical technique for sectioning lobster  
epidermis and staining for enzyme activity. I have been trying to  
find an ideal freezing temperature for the cryostat. The technique I  
am adapting was originally developed for scyphozoans, and calls for  
freezing temp of 30 C. This appears to be too cold, as the tissue  
just crumbles the blade crosses it. I have tried 20 C, and this  
appears to be better, but I was wondering if anyone has experience  
with freezing arthropod or crustacean tissues, and might have a  
suggested temp.

Thanks,
Norman



Begin forwarded message:

> From: mwstarbu@mdanderson.org
> Date: August 1, 2007 2:10:40 PM EDT
> To: Histonet@pathology.swmed.edu, histonet- 
> bounces@lists.utsouthwestern.edu
> Subject: Re: [Histonet] Thin sectioning bone in PMMA
>
> We apply a 50% butyl glycol, 35% ethanol solution to the  
> sections.   The
> application is as follows.  We apply a liberal amount with a small  
> paint
> brush and let dry 5 -10 minutes under a fume hood.  Apply again and  
> use
> downward pressure with the brush to remove wrinkles. During the 2nd
> application you will notice the sections are much more pliable  
> enableing
> you to easily remove large wrinkles.  We then cover the slides with
> plastic strips, press and dry at 55 degrees C overnnight
>
> Mike
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