[Histonet] air drying special stain slides rather than dehydrate and clear

[Rene J Buesa]

Hazel:
The so called "drying artifact" that appears as a "darker granulation", like sand grains, is caused NOT by drying the sections, but by drying them incompletely. IF there is any remnant of water within or between the cells, that water will refract the light using to observe the sections and will appear as "dark granules". It is the same effect as when you observe rain falling in the horizon: even when the water is transparent, you will see the falling rain as a darker film, almost black, also because of light refraction.
If you completely eliminate the water, that artifact will not take place.
Under separate cover I am sending something about this I published.
By the way, I was taught the same when I started to study histology during my pre-medical studies in 1952 (so do not talk about age, I am for sure older than you are!). Keep your brain young, it does not matter if your body ages, your brain has to remain young!
René J.


________________________________
From: "Horn, Hazel V" <HornHV <@t> archildrens.org>
To: 'Amos Brooks' <amosbrooks <@t> gmail.com>; "histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu> 
Sent: Wednesday, September 12, 2012 8:57 AM
Subject: RE: [Histonet] air drying special stain slides rather than dehydrate and clear

I suppose my age will show here but I was always taught to NEVER let slides dry out unless the procedure indicated such.  Is there no drying artifact when you let these slide dry before coverslipping?

Hazel Horn
Supervisor of Histology/Autopsy/Transcription
Anatomic Pathology
Arkansas Children's Hospital
1 Children's Way | Slot 820| Little Rock, AR 72202
501.364.4240 direct | 501.364.1302 office | 501.364.1241 fax
hornhv <@t> archildrens.org
archildrens.org




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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Amos Brooks
Sent: Tuesday, September 11, 2012 7:31 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] air drying special stain slides rather than dehydrate and clear

Hi,
    My choice to air dry rather than dehydrate in ETOH & xylene is based on the stain rather than the spooky xylene hazard boogyman. Yes, not using xylene if it is not really needed is not a bad idea, but the main reason I air dry some stains is the alcohols remove some of the stains. Ever have a beautiful Luxol Fast Blue bleach out on you? The most exasperating thing in the world!
    Generally stains that end in water can easily be air dried. Something alcoholic like eosin or Movat's Pentachrome ending in alcoholic saffron might as well be finished traditionally. I air dry any stain that is counterstained in Nuclear Fast Red, Light Green, Methyl Green. I have air dried IHCs with no ill effects too. Don't try it with fluorescents though, that would be bad ... and pointless.
    I don't put them in an oven. I set them at the front of the fume hood and go do something else for a few minutes. If I want to rush it I close the sash to increase the flow rate for a bit. (Of course it is opened back up right after so the draft works properly.) Amos


On Tue, Sep 11, 2012 at 11:09 AM, <histonet-request <@t> lists.utsouthwestern.edu
> wrote:

> 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
>
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