[Histonet] Pap stain without xylene

[Mike Toole]

Thank you Beth,

It’s good to know that someone else had tried this and had very good results. The method you suggest is very much in line with the recommendations from René. He did recommend using a drying oven at 60°C to help ensure complete removal of any water or alcohol. And, that absolute dryness was a requirement for coverslipping without artifact such as the appearance of sand like grains or cornflakes.

Just to reiterate, was the field method performed at ambient temperature without the aid of a drying oven? And, just a thought, I suppose if the lack of electricity was an issue in a field setting, that perhaps a solar oven made with plywood and glass could be used to elevate temperature for drying.

Do you  know if altering the method for final clearing would require validation?

Mike

From: Beth Cox [mailto:bethcoxx at gmail.com]
Sent: Wednesday, May 25, 2016 12:56 PM
To: Mike Toole; Histonet
Subject: [EXTERNAL] Re: Pap stain without xylene


Mike,



As an alternative for your discussion:



I work on medical mission trips where I run Paps in a field setting.  For the past 4 years, we have not had access to xylene, and we have done the following:



1. Remove slides from the last alcohol after staining

2. Allow to air dry completely (generally only takes a few minutes)

3. Use standard mounting media to coverslip them dry.



This works beautifully.  Microscopically they look the same as xylene cleared slides.  The only caveat is that they must dry completely.



--

BETH COX, HTL/SCT(ASCP)QIHC

AP Consultant | Pathology Solutions, Inc | (810) 240-2190 | bethcoxx at gmail.com<mailto:bethcoxx at gmail.com>





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Message: 6

Date: Tue, 24 May 2016 17:08:51 -0500

From: Mike Toole <mtoole at dcol.net><mailto:mtoole at dcol.net>

To: "histonet at lists.utsouthwestern.edu"<mailto:histonet at lists.utsouthwestern.edu>

        <histonet at lists.utsouthwestern.edu><mailto:histonet at lists.utsouthwestern.edu>

Subject: [Histonet] Pap stain without xylene

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Ren? 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