[Histonet] Porcessing FFPE tissue without alcohol??

Rene J Buesa rjbuesa at yahoo.com
Sat Mar 26 10:35:36 CDT 2016


Sudan Black reacts only with protein-combined fats.René 

    On Saturday, March 26, 2016 11:20 AM, Joanna <jobaluyut at gmail.com> wrote:
 

 How about Sudan Black stain? 

> On Mar 26, 2016, at 4:32 AM, Rene J Buesa via Histonet <histonet at lists.utsouthwestern.edu> wrote:
> 
> The only problem I see is that the fat will be preserved, as you wrote, as a black osmium oxidate but you will not be able to use any "standard" fat stain; otherwise it will work.René 
> 
>    On Friday, March 25, 2016 2:41 PM, "Hobbs, Carl via Histonet" <histonet at lists.utsouthwestern.edu> wrote:
> 
> 
> 
> 
> Fix the tissue in Formalin, wash well in dw, then place very small pieces into Osmium tetroxide solution ( std soln for TEM post-fixation)
> Processing to Pwax as usual.
> Basically, you will see lipids as black ( oxidised osmium)
> That's the only way to demonstrate solvent- soluble lipids, using Pwax processing.
> Sure, there are caveats but, in the main...it will be Ok, imho.
> I invite comments as I may be doing exactly this very soon, to count myelinated nerve fibres in a sciatic nerve.
> 
> 
> 
>  
> 
> Carl Hobbs FIBMS 
> Histology and Imaging Manager 
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