[Histonet] RE: air drying special stain slides rather than

Tue Sep 11 12:47:27 CDT 2012

We use tape coverslipping. Some techs dip the slides in the xylene on stainer before adding them to the coverslipper and some just use the amount of xylene coming out of the drip on the coverslipper. Either works fine and we have had no problems.

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Diana McCaig
Sent: Tuesday, September 11, 2012 10:23 AM
To: E. Wayne Johnson; Rene J Buesa
Cc: histonet <@t> lists.utsouthwestern.edu; Mayer, Toysha N
Subject: RE: [Histonet] RE: air drying special stain slides rather than

Would this work for auto cover slipping  (tape film)if they were set in the xylene reservoir prior to cover slipping?

Diana

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of E. Wayne Johnson
Sent: September-11-12 1:15 PM
To: Rene J Buesa
Cc: 'histonet <@t> lists.utsouthwestern.edu'; Mayer, Toysha N
Subject: Re: [Histonet] RE: air drying special stain slides rather than

I am convinced to give it a try because I also have trouble will the loss of some stains in dehydration.
I was concerned that the slides would not clear well after oven dehydration.  I will see how it works for me.

I can see clearly how going from counterstain to oven will save much hassle with xylene and alcohols as well as not washing out some special stains.  I have tried some of the isopropyl alcohol and acetone dehydration called for in some of the stain procedures and it would be great if the slides could just be popped into the oven.

What mounting medium are you using?  Does it matter?  I am a bit worried about penetration of the mountant into the tissue section if there is no xylene in the tissue.  Will neutral balsam still work ok?

Rene:  if you have a link to the paper you talked about on eliminating xylene, I am interested.  Xylene is becoming more and more of an issue and a pain for us.

EWJohnson
Enruikang Ag Tech
Beijing.

On 9/12/2012 12:01 AM, Rene J Buesa wrote:
> Toysha:
> Perhaps you have not oven dried stained slides before, and that explains some of your comments, like:
> 1- if the stained slides are completely dried, the "miscibility" you
> point out is not an issues, because there is nothing to mix with;
> 2- if you dehydrate → clear the stained sections that will take about
> 15 minutes per group of up to 25 slides, or even more depending on the
> protocol used in your automated stainer, but if your group of slides
> in their rack are placed in an oven at 60ºC for 5 minutes it will just
> that, 5 minutes reducing the usual TAT for each staining procedure;
> 3- any oven can accommodate more than 100 stained slides in their
> racks and the TAT is shortened by oven drying, no matter how many
> slides you are working with;
> 4- I really do not know where you can find that "extreme heat" can
> affect the tissue sections. All tissue sections are fixed → processed
> → dried (usually at the same 60ºC before staining) → stained and an
> additional step at 60ºC to dry before cover-slipping is just that, an
> additional step at 60ºC
> 5- The so called "Lean" technologies do not refer to staining only,
> they have to do with the whole work-flow and an additional drying step
> at 60ºC cannot affect in a negative way to the work-flow
> 6- after staining you will oven dry the sections.
> I think you should try the method instead.
> René J.
>
>
> ________________________________
> From: "Mayer,Toysha N"<TNMayer <@t> mdanderson.org>
> To:
> "'histonet <@t> lists.utsouthwestern.edu'"<histonet <@t> lists.utsouthwestern.ed
> u>
> Sent: Tuesday, September 11, 2012 11:41 AM
> Subject: [Histonet] RE: air drying special stain slides rather than
>
>
> Ooh, great question for my students next semester.
> Your answer is the counterstain, some counterstains may require dehydration after rinsing, or some may not. Adjusting the times of the counterstain is not the issue as much as  the solvent of the counterstain.
>
> Rene, while I do acknowledge that the xylene may/will cause hazards, we must think of the miscibility of the clearant and the dehydrant, as well as the amount of time involved.  The amount of time involved to blot and air dry the slides will affect the TAT for the specimen.  5 min may be ok if you have a small amount of slides, but with a larger number of slides, it will be considerably more than 5.  Also Lean methodologies would not apply in that case. With automation, the extreme heat involved with a stain dryer may affect the tissue on the slide.
>
> There are some stains that can be blotted, cleared and coverslipped, but using the alcohol to remove excess water and counter stain is better in my opinion.
>
>
> Toysha N. Mayer, MBA, HT (ASCP)
> Instructor, Education Coordinator
> Program in Histotechnology
> School of Health Professions
> MD Anderson Cancer Center
> (713) 563-3481
> tnmayer <@t> mdanderson.org
>
>
>
>
> Message: 16
> Date: Tue, 11 Sep 2012 10:32:08 -0400
> From: "Diana McCaig"<dmccaig <@t> ckha.on.ca>
> Subject: [Histonet] air drying special stain slides rather than
>      dehydrate    and clear
> To:<histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
>      <DCFD9E6A390E294AAF3A2561CD32E5C417A90529 <@t> ckhamail1.ckha.on.ca>
> Content-Type: text/plain;    charset="us-ascii"
>
> I was hoping to get information on why special stains are dehydrated, cleared and mounted vs allowing them to be blotted dry, air dried then coverslip.
>
>
>
> Every procedure I have ever encountered always indicates to dehydrate and clear but I have heard where some labs are blotting the slides , allowing to air dry (probably not set standard time) and dipped in xylene prior to cover slipping.  Reason given is that the counterstain gets washed out.  Wouldn't adjusting the times be a better resolution.
>
>
>
> I understand residual water could be present and cause long term issues on storage but wanted some other opinions on this process.
>
>
>
> Diana
>
>
>
> ------------------------------
>
> Message: 17
> Date: Tue, 11 Sep 2012 07:52:05 -0700 (PDT)
> From: Rene J Buesa<rjbuesa <@t> yahoo.com>
> Subject: Re: [Histonet] air drying special stain slides rather than
>      dehydrate    and clear
> To: Diana McCaig<dmccaig <@t> ckha.on.ca>,
>      "histonet <@t> lists.utsouthwestern.edu"
>      <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
>      <1347375125.72189.YahooMailNeo <@t> web121405.mail.ne1.yahoo.com>
> Content-Type: text/plain; charset=iso-8859-1
>
> Diana:
> The most simple answer to your question is: "Because that is the way it has been done for more than 150 years".
> The second question would be: "Is it necessary?" and the short answer to this question is: NO!!!
> As a matter of fact, one of the steps I have developed to totally eliminate xylene from the histology lab refers to the "clearing" of stained sections, not only "special stains" (the so called HC and IHC) but the routine as well (the H&E).
> Now, the "secret" to a successful drying of the stained slides is NOT to let them air dry because that will take not only too much time, but you can never be sure if the section is completely dry and if you add the mounting medium to a not completely dried section, you will have transparency problems.
> The correct way of doing that is by drying the stained sections during 5 minutes at 60?C in an oven.
> Under separate cover I am sending you something I published about your question and other aspects of how to completely eliminate xylene from ALL steps in the histology laboratory.
> Ren? J.
>
>
> ________________________________
> From: Diana McCaig<dmccaig <@t> ckha.on.ca>
> To: histonet <@t> lists.utsouthwestern.edu
> Sent: Tuesday, September 11, 2012 10:32 AM
> Subject: [Histonet] air drying special stain slides rather than
> dehydrate and clear
>
> I was hoping to get information on why special stains are dehydrated, cleared and mounted vs allowing them to be blotted dry, air dried then coverslip.
>
>
>
> Every procedure I have ever encountered always indicates to dehydrate and clear but I have heard where some labs are blotting the slides , allowing to air dry (probably not set standard time) and dipped in xylene prior to cover slipping.? Reason given is that the counterstain gets washed out.? Wouldn't adjusting the times be a better resolution.
>
>
>
> I understand residual water could be present and cause long term issues on storage but wanted some other opinions on this process.
>
>
>
> Diana
>
>
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