[Histonet] vibratome sectioning of brain tissues
A colleague of me works with rat brains and tries to prepare vibratome section but with hardly no result. =20
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
-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 37║
-during the next 24 H hemispsheres were infiltrated with gelatin, 25% w/w at 37║
-after coagulation (approx 30 min) at 2-10 ║ gelatin blocks were further fixed during 4
days in 4% formaline (2-10 ║).
A (new) vibratome, HM 650 V, was used for sectioning.
Intended section thickness: 35 Ám
The buffertray was filled with PBS and crushed ice
She have tried many different combinations of the following settings:
-knife angle: 10-40 ║ (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!!!
TNO Quality of Life
Toxicology and Applied Pharmacology
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