[Histonet] Embedding paraffin for brain tissue?

[Pirici Daniel]

Hi there,
 
1. Have you changed anything in the processing and embedding processes themselves (dehydration, clearing agents and times, etc)? Friable sections could also come from extended times in the clearing agent...
 
2. There is also a section transfer system with a waterfall. We have a microtome which has both systems to collect the ribbons (classical and with the waterfall). There is a huge difference between them, it is much easier to cut on the water-based system. We can cut even 100 seriate sections without losing even 1 section!!! And there are also Peltier-based cooling block clamps that help keeping your paraffin bloc cold during the cutting!!! There are many companies offering these options...
 
But since you are experiencing a sudden change in the quality of the ribbons with the same paraffin initially, I would seriously consider first some change in the embedding protocol...
 
I hope this might help!
 
A HAPPY NEW YEAR TO ALL HISTONETT-ERS!!!!!

________________________________________________________________________________________________ 
Pirici Nicolae Daniel, MD, PhD 
Department of Histology
University of Medicine and Pharmacy Craiova
Petru Rares Street 2, 200349
Craiova, Dolj
Romania
 

________________________________
 From: Maria Mejia <mbmphoto <@t> gmail.com>
To: Tim Wheelock <twheelock <@t> mclean.harvard.edu> 
Cc: "histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu> 
Sent: Thursday, January 2, 2014 8:04 AM
Subject: Re: [Histonet] Embedding paraffin for brain tissue?
  

Hello,

I too have been experiencing the same difficulties cutting brain samples, I've tried 3 different
types of paraffins - all the the same rolling of sections, some lines & tearing.  It's not my
microtome, because I've had it checked & it went through a recent general maintenance.
I've tried several angle changes, used different brands of low profile coated blades &
carefully watch the temperature of the tissue blocks.

I'm also feeling very frustrated for not being able to cut good brain sections.  Any assistance
anyone can provide will be greatly appreciated.

Maria Mejia
UCSF 
Department of Neurology
San Francisco, CA


On Dec 31, 2013, at 8:41 AM, Tim Wheelock wrote:

> Hi Everyone:
> 
> In November, there was a discussion concerning different types of embedding paraffin.
> Do people find that a certain kind of wax is preferable for embedding brain tissue, or doesn't it matter?
> 
> I also use Surgipath EM-400, but have decided to try another brand.
> I have been using this paraffin for about 25 years.
> It used to cut like butter, with beautiful ribbons and no lines.
> I would put the blocks on ice, wait 2 hours, and then cut 7-8 cases (about 190 slides) per day easily.
> Now I am struggling to cut 4-5 cases.
> I am experiencing a lot of lines in my sections and some rolling and tearing of parts of the sections.
> This is affecting my lab's productivity, the quality of my sections, and is a source of constant frustration.
> 
> I am cutting with Surgipath high profile Teflon coated blades.
> When I tried Thermo-Fisher HP-35 Ultra blades, at first they helped, but soon I experienced rolling of the sections.
> I have tried changing the angle of the blade, to no avail.
> So, I thought that I would try a change in the embedding paraffin.
> 
> Thanks for any suggestions that you may have,
> 
> Tim Wheelock
> Harvard Brain Bank
> McLean Hospital
> Belmont, MA
> 
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