[Histonet] Vibratom operation - HELP
Olek Michalski
olek.michalski <@t> nencki.gov.pl
Thu Mar 6 18:26:23 CST 2008
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
chamber.
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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