RE: [Histonet] mouse heart chatter upon sectioning...

From:Laurie Reilly



Dina,

Rodent tissues are notorious for becoming brittle, even with the gentlest
processing. 

Try rehydrating the tissues a little in the paraffin block.
After you trim the face of the paraffin block, float it face down on your
waterbath for 30 seconds. 
Dry the face of the block, put it back on ice to cool, and then try
sectioning. You should be able to get 5 or 6 good sections before it starts
to shatter again.

This is a fact of Histo life that I have lived with for 30 years and I would
love to hear a scientific reason why some tissues, particularly from
rodents, small birds and reptiles give this trouble.

Can anyone help with an explanation?

Regards,   Laurie.

Mr. Laurie REILLY
Histopathology
School of  Veterinary and Biomedical Sciences
James Cook University
Townsville  Qld.  4811
Australia.

Phone 07 4781 4468

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gaupp, Dina
D 
Sent: Thursday, 5 June 2008 9:16 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] mouse heart chatter upon sectioning...

Whenever I process (whole)mouse hearts, the tissue chatters upon sectioning.
The tissue is usually fixed overnight in 10%nbf at 4C before processed.  I
tried different processing times.  The last time I processed hearts, I
processed them for 5 hours total no vaccum.  Should I decrease the
processing time even lower than that?  I am afraid of underprocessing.  I
have to section the entire heart from top to bottom, so I really need some
advice.
 
(open system) Processor Set Up:
1 - fix (if tissue fixed overnight, skip #1 & #2 stations)
2 - fix
3 - 70%
4 - 80%
5 & 6 - 95%
7 & 8 - 100%
9 & 10- Xylene Sub. - Clear-rite
11 & 12-Paraffin stations
 
Thanks,
Dina
dgaupp@tulane.edu  
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