[Histonet] vibratome sectioning of brain tissues
Bruijntjes, J.P.
bruyntjes <@t> voeding.tno.nl
Fri Mar 10 10:19:38 CST 2006
Hi all
A colleague of me works with rat brains and tries to prepare vibratome section but with hardly no result.
Main problem is thin/thick sections.
This is what she has done with the tissues ect:
Brain tissue: perfused with and fixed pre-embedding in 4% Formaline
Embedding procedure:
-during 24 H hemispheres were washed in running tap water
-during the following 24 H hemispheres were infiltrated with gelatin (from bovine skin
type B) solution, 12.5% w/w at 37º
-during the next 24 H hemispsheres were infiltrated with gelatin, 25% w/w at 37º
-after coagulation (approx 30 min) at 2-10 º gelatin blocks were further fixed during 4
days in 4% formaline (2-10 º).
A (new) vibratome, HM 650 V, was used for sectioning.
Intended section thickness: 35 µm
The buffertray was filled with PBS and crushed ice
She have tried many different combinations of the following settings:
-knife angle: 10-40 º (approx)
-frequency: 30-100 Hz
-amplitude: 0.3-1.2 mm
-speed: 1.5-5.0 mm/s
She is running out of ideas. Suggestions are very welcome!!!
Joost Bruijntjes
TNO Quality of Life
Toxicology and Applied Pharmacology
Zeist
Holland
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