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

Wed Sep 12 19:32:11 CDT 2012

We have routinely dried slides prior to coverslipping. We have found that ethanol or acetone rinsing after staining shortens the drying time (in fact we have used acetone to dehydrate MGG and DiffQuik stained smears, which must be kept away from alcohols, prior to coverslipping).

Our automatic coverslipper uses a very runny xylene based mountant so we do not need to rinse in xylene prior to coverslipping.

One paper we did describes the detergent de-waxing aspect and a study currently in preparation applies the technique to fungal staining:

Henwood A (2012) "The application of heated detergent dewaxing and rehydration to immunohistochemistry" Biotechnic & Histochemistry 87(1): 46-50.  

It will depend on the staining method used as to whether you can use alcohol, acetone or heat-assisted drying prior to coverslipping, but, dare I say, nearly all stains can be treated thus.

Whoops, I forgot about the Oil Red O stains for fats, Oh well I did say "nearly all"!!

Regards 
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Laboratory Manager & Senior Scientist 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Mayer,Toysha N
Sent: Wednesday, 12 September 2012 1:42 AM
To: 'histonet <@t> lists.utsouthwestern.edu'
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

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

*********************************************************************************
This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender.

Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead

This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses.
*********************************************************************************