[Histonet] Stamp staining in paraffin and smear samples

[Julio Benavides]

Hi There,

Is anyone using this technique to stain chlamydia in placental samples  
(working with sheep here)? I was looking for a working technique but  
it looks like it is not easy to find.

Thanks for your thoughts!

Julio


Rene J Buesa via Histonet <histonet at lists.utsouthwestern.edu> escribió:

> Hi Mike:The steps you desrcibe are wrong.After you finish staining  
> your PAP smear, just wash them in the last ethanol → oven dry at  
> 60ºC for 5 minutes or as required if the smear is too thick and when  
> completely dried → coverslip.This final drying has to take place at  
> temperatures above room temp. because you have to be absolutely sure  
> the smear (or stained tissue section)  is absolutely dried before  
> applying the mounting medium → coverslip.The steps you write about  
> 2-propanol and mineral oil are those the tissues have to go through  
> before paraffin infiltration. René
>
>     On Tuesday, May 24, 2016 6:36 PM, Mike Toole via Histonet  
> <histonet at lists.utsouthwestern.edu> wrote:
>
>
>  René and other Histonetters,
>
>
> After reading the paper:
>
>
>
> Buesa RJ, Peshkov, MV. Histology without xylene. Ann Diagn Pathol.  
> 2009 Aug;13(4):246-56. Epub 2009 Feb 5.
>
>
>
> It appears that xylene in the final clearing steps is replaced with  
> isopropanol and mineral oil as follows:
>
> ·        5:1        isopropanol to mineral oil            50°C
>
> ·        2:1        isopropanol to mineral oil            50°C
>
> ·        Undiluted mineral oil                                    50°C
>
> ·        Drying oven 5 minutes                                  60°C
>
> ·        Coverslip
>
>
>
> Do you feel these clearing steps be applied to the pap stain in  
> order to eliminate xylene?  If so, can it be done at room temperature?
>
> Thanks,
> Mike
>
> Mike Toole, BS, CT(ASCP)CM
>
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