[Histonet] need your kind help preparing sections of a whole rabbit eye

[Frentz <@t> ACTO <@t> home]

Dear all,

 

A common problem after the fixation of a rabbit's eye in 3.7% Formalin/PBS
solution is to cut though the middle of the whole eye with out loosing the
structural integrity, meaning ripping out the lens.

I now decided to deep freeze the globe in N2 and cut it using a micro band
saw, hope it will work.

 

But the delicate part starts after that.

 

Dehydrating the bisected tissue for paraffin embedding causes hardening of
the lens. This makes the sample inappropriate for cutting causing again loss
of the lens out of its holding apparatus in the paraffin block.

 

My idea now was to embed the tissue in a water containing material like agar
or gelatine. Because I do not have lots of experience doing histological
preparations, I searched the web and luckily found the histonet, obviously
with lots of professionals.

 

Searching the site gave me the idea, that gelatine embedding is usually done
preparing samples for the vibratome which we do not have in the lab.

 

Do you think it can be used for slide or rotation microtoms and how thin
will the slices be at best?

 

Can you give me useful recipes and a working procedure to solve the problem
because I need to observe the whole inner eyeball following impact force to
the cornea from different distances?

 

Thanks in advance!!

 

Markus   

 

 

-------------------------------------------------------------------
  Markus Frentz

  Department for Ophthalmology

  Research Lab

  Pauwelsstrasse 30

  52070 Aachen

 

  Germany

  Fon (Lab): +49/241/80-88224
  Fax (Lab): +49/241/80-3388224

 
  web:  <http://www.augenklinik.ukaachen.de> www.augenklinik.ukaachen.de