[Histonet] Dead-End system & BMA/MMA embedded tissue

[Sven van Eijl]

Dear Histonetters,

This will be my first posting to this list, and I am by no means a real histologists, so bear with me please...
We are having a problem using Promega's Dead-End colorimetric TUNEL kit on plastic embedded lungs.
In the manual are only guidelines for formalin fixed and parrafin embedded or frozen tissue. We fixed the lungs using Carnoy's and embedded in BMA/MMA. I have had some tips from Promega about modifications to the protocol to account for the use of other fixatives than formalin, but am wondering if it is the fixative or the plastic that is causing the problems. We get a nice brown staining of all nuclei at all times, also in the negative controls. It is not a question of general high background, the rest of the tissue is clear. Any suggestions are highly appreciated.

Thanks,
Sven


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T.J.A.van Eijl, MSc.
Dept. Pharmacology and Pathophysiology               
Utrecht Institute for Pharmaceutical Sciences        
Utrecht University
Utrecht
The Netherlands                                   

F.A.F.C. Wentgebouw
Sorbonnelaan 16
3584 CA Utrecht
Room W146               

Tel:  +31(0)30 253 1215
Fax: +31(0)30 253 7420

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"Many a man fails to become a thinker for the sole reason that his memory is too good"
Nietzsche
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