[Histonet] Xylene Substitutes

[Rene J Buesa]

Donna:
Please go to the web site I indicated in my previous answer and you find detailed answer.
After you stain the sections (regardless of the procedure) you dry them in an oven at 60ºC for 5 mins. and they are completely dry (ANATHEMA!!!!!!! to most) you just place them in your automated coverslipper with the same mounting medium you are using now. You need to change NOTHING.
Try it!
René J.


________________________________
From: "Dingersoll <@t> aplaboratories.com" <Dingersoll <@t> aplaboratories.com>
To: Rene J Buesa <rjbuesa <@t> yahoo.com>; "Yaskovich, Ruth A (NIH/NIDCR) [E]" <ryaskovich <@t> dir.nidcr.nih.gov>; "'Histonet <@t> lists.utsouthwestern.edu'" <Histonet <@t> lists.utsouthwestern.edu> 
Sent: Wednesday, August 21, 2013 12:20 PM
Subject: RE: [Histonet] Xylene Substitutes



Rene,

How do you recommend cover slipping with out xylene?  We use a tape cover slip instrument that requires xylene.  If you are using glass coverslips what kind of adhesive is used?  I am very interested in eliminating xylene as much as possible, but we are a high volume lab and many of the automated instruments (stainers, coverslippers) require xylene.  Thanks in advance for your feedback.

Donna S. Ingersoll, B.S., HTL, CT(ASCP)
Laboratory Manager
A P Laboratories, LLC
2090 Executive Hall Rd Suite 165
Charleston, SC 29407
843-300-3001 X 202
843-300-3003 (fax)
dingersoll <@t> aplaboratories.com

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-------- Original Message --------
>Subject: Re: [Histonet] Xylene Substitutes
>From: Rene J Buesa <rjbuesa <@t> yahoo.com>
>Date: Wed, August 21, 2013 12:06 pm
>To: "Yaskovich, Ruth A (NIH/NIDCR) [E]" <ryaskovich <@t> dir.nidcr.nih.gov>,
>"'Histonet <@t> lists.utsouthwestern.edu'"
><Histonet <@t> lists.utsouthwestern.edu>
>
>Ruth:
>I all reality none is good. Both alkanes or D-Limonene are either as dangerous or more than xylene, and all perform below the xylene standard.
>Any one requires modifications of all procedures, they seldom dewax adequately and all constitute a processing "compromise". Many cannot de recycled.
>Please go to http://www.histosearch.com/rene.html and read about xylene substitution.
> 
>The optimal solution to eliminate xylene consists on:
>1- dehydrate with 2-propanol (or "pure" isopropyl alcohol or IPA)
>2- do not use any ante-medium and go from IPA directly to paraffin wax or ideally to mineral oil:IPA at 1:1
>3- infiltrate as usual
>4- dewax sections with 2%aq. sol. of dishwater soap at 90ºC for 1 min twice
>5- stain as usual
>6- dehydrate stained sections in a convection oven at 60ºC for 5 minutes
>7- coverslip as usual.
>As you can see xylene is nowhere to be found in this sequence that produce optimal quality slides.
>Try it!
>René J.
>
>
>________________________________
>From: "Yaskovich, Ruth A (NIH/NIDCR) [E]" <ryaskovich <@t> dir.nidcr.nih.gov>
>To: "'Histonet <@t> lists.utsouthwestern.edu'" <Histonet <@t> lists.utsouthwestern.edu> 
>Sent: Wednesday, August 21, 2013 11:18 AM
>Subject: [Histonet] Xylene Substitutes
>
>
>I know this question has been asked before but what's the best and safest Xylene Substitute?
>Ruth Yaskovich
>N.I.H. N.I.D.C.R.
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