[Histonet] Re: Histonet Digest, Vol 73, Issue 39

[rgrow <@t> bmnet.com]

R C,
In past years, I've section many primate brains in normal  size sections
but not whole, so this may/may not help.
If it was fixation you would not be able to get a complete section in the
first place, so your fixation should not be a problem.  If it was the
adaptor on your microtome you would have other sectioning problems: wobble,
thick/thin, washboard, curved sections, uneven thickness, etc.
A couple of suggestions that might help.  The suggestions should work
regardless of section thickness.

1.  Try initially laying your ribbon on a dish of COLD water.  With fine
forceps, (chilled too) gently stretch out your section. When you are
satisfied pick your section up from the cold and transfer it to the warm
water to complete the process.  Lengthens your cutting time, but works well
if you have the time.

2.  Be sure to use good qualilty high profile blades, if your
stage/faceplate will accomodate them.  Surgipath has good ones.  For large
sections they are more stable.

3.  I did use the Surgipath Infiltration Medium for the processor.  And,
yes, it does make a difference.  Contact your Surgipath Sales Rep and
request  "Infiltration Medium" to set up in your processor.  You might be
able to get enough for a trial run.  Continue to use your "Type R" for
embedding.

You may need a combination of these things to get the quality you are
looking for.

Let me know if I can help further.

Happy New Year to ALL  Histonetters!

Renee Grow, BA., HT (ASCP)
rgrow <@t> bmnet.com
Histology Supervisor
Blount Memorial Hospital
907 E. Lamar Alexander Pkwy.
Maryville, TN  37804-5016
(865) 977-4744
(865) 977-5766 Fax

You wrote:

Message: 4
Date: Tue, 29 Dec 2009 14:16:04 -0800
From: R C <ruebenjcarter <@t> gmail.com>
Subject: [Histonet] 2x3 microtomy: Adapter vs Sliding Microtome
To: histonet <@t> lists.utsouthwestern.edu
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		 <2a926e3f0912291416u723bd5d4ya6b93e199b4a3d0 <@t> mail.gmail.com>
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Hi. I'm working on a protocol for cutting monkey brain sections to be
mounted on 2x3 slides. I've read about utilizing a sliding microtome but in
short, have decided to use the 2x3 adapter for a standard Microm microtome.
During microtomy I've noticed many wrinkles in the sections, particularly
within the folds of the cerebellum. The wrinkles worsen as the sections
float in the water bath (temp=38).

In troubleshooting, a co-worker suggests inadequate fixation. I on the
other
hand believe that the wrinkles relate to the Type R paraffin, which
contains
polymers as well as the use of the adapter versus the sliding microtome.

Can anyone offer any first hand experience/guidance?