[Histonet] Vibratom operation - HELP

From:"Olek Michalski"

Dear Histonetters,

I am in major trouble. I am slicing Golgi-Cox impreganted brains into  
120um sections. I used to embed tissue in 4% agar for slicing but it  
blocks cracked and the tissue was not cut very well. My coleague told me  
she prefer 30% gelatin for blocks, so I tried it and the sections are  
shattered. There are actually tiny stripes of tissue kept together by the  
gelatin block (or sometimes even not).
I have to admit the blocks are really hard - they are composed of 30% w/w  
gelatin. The tissue was impreganted with Golgi-Cox solution (chromates and  
sublimate) for two weeks, then transferred to 30% sucrose for about a  
month then soaked with 10% gelatin (2 days in 37 deg.) and 30% gelatin (1  
day about 50 deg.). I normally use 6% sucrose to fill the sectioning  
Could anybody help me, please?

Yours sincerely
Olek Michalski
        Laboratory of Neurobiology
       of Development and Evolution

  Nencki Institute of Experimental Biology
  ul. Pasteura 3, 02-093 Warszawa,  Poland
  Tel. +48 22 5892268,  Fax +48 22 8225342

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