[Histonet] Pap stain without xylene

[Rene J Buesa]

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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