[Histonet] vibratome sectioning of brain tissues

[Bruijntjes, J.P.]

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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