Re: [Histonet] Endothelial Cell Border stains

From:Rene J Buesa

  Bensley & Bensley  (Anat.Rec.,vol 64 pp 46,1935) descibe a method with silver chloride dichlorfluoresceinate. It requires to inject the animal intravenously with 0.8% aq. dichlorofluorescein until the animal becomes quite yellow. Kill the animal and immerse the targeted tissue in 10% aq. silver nitrate. Fix in 10% neutral formalin, dehydrate in ethanol, clear in iso-safrol, mount in balsam. The endothelial cells will be outlined in pink. Exposed to light silver becomes park brown to black with defined outlines. Silver inconsistencies are usually due to procedural inconsistencies.
  Hope this will help!
René J.

"Bond, Andrew"  wrote:
I am trying to look at changes in endothelial cell morphology in the
rabbit thoracic aorta. I have managed to get Hautchen preparations to
work fairly well, and now need to stain endothelial cell borders. Only
problem is I am trying to carry out automated image analysis therefore
need a very uniform and specific fluorescent stain, that leaves very
clear lines without any cytoplasm/nuclear staining. I've been told
silver nitrate staining would work but is very inconsistent. Does
anyone have any suggestions?

Andrew Bond
Imperial College London, UK
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