[Histonet] "FREEZY" spray

Wed Apr 8 00:38:00 CDT 2009

Sorry, again, I'm confused... are you responding to me? I'm not the original poster... I never stated I had trouble with cracking, because, well, I don't. I'm Bernie Taupin, the King of Cryomicrotomy, Esq.

________________________________
From: Akemi Allison-Tacha <akemiat3377 <@t> yahoo.com>
To: Jennifer MacDonald <JMacDonald <@t> mtsac.edu>; Ingles Claire <CIngles <@t> uwhealth.org>; Bernie Taupin <bernietaupin <@t> ymail.com>
Cc: Histonet <@t> lists.utsouthwestern.edu; histonet-bounces <@t> lists.utsouthwestern.edu
Sent: Wednesday, April 8, 2009 1:27:35 AM
Subject: Re: [Histonet] "FREEZY" spray

Sorry, I too noticed that the text looked weird then I did a cut and paste.  I keep all the Data Sheets I created in my files, and this was just a portion of the Notes from the original Data sheet.  I thought it might be helpful because you stated you had difficulty with cracking. 

You did not mention using isopentane.  Some people immerse the tissue straight into liquid nitrogen, which could cause freezing artifacts.

Akemi Allison-Tacha BS, HT (ASCP) HTL
Histology Manager
Associated Pathology Medical Group Laboratories
105A Cooper Court, Los Gatos, CA 95032 
Cell: (425) 941-4287
E-Mail: akemiat3377 <@t> yahoo.com

--- On Tue, 4/7/09, Bernie Taupin <bernietaupin <@t> ymail.com> wrote:

From: Bernie Taupin <bernietaupin <@t> ymail.com>
Subject: Re: [Histonet] "FREEZY" spray
To: "Akemi Allison-Tacha" <akemiat3377 <@t> yahoo.com>, "Jennifer MacDonald" <JMacDonald <@t> mtsac.edu>, "Ingles Claire" <CIngles <@t> uwhealth.org>
Cc: Histonet <@t> lists.utsouthwestern.edu, histonet-bounces <@t> lists.utsouthwestern.edu
Date: Tuesday, April 7, 2009, 10:11 PM

I use isopentane suspended in liquid nitrogen, too. 

sorry if this sounds curt, but due to your cut-and-pasting, im not entirely sure what point youre trying to get at...?

________________________________
From: Akemi Allison-Tacha <akemiat3377 <@t> yahoo..com>
To: Jennifer MacDonald <JMacDonald <@t> mtsac.edu>; Ingles Claire <CIngles <@t> uwhealth.org>; Bernie Taupin <bernietaupin <@t> ymail.com>
Cc: Histonet <@t> lists.utsouthwestern.edu; histonet-bounces <@t> lists.utsouthwestern.edu
Sent: Wednesday, April 8, 2009 1:03:04 AM
Subject: Re: [Histonet] "FREEZY" spray

Bernie,

I used to use the method below for FS on muscles at Emanual Hospital in Portland OR. Since we had such wonderful success, we incorporated this method for all FS.  

Years later, I developed the "MATSSE (pH3.4) 1-Step Trichrome Stain Kit" (Modified Trichrome Method for Muscle Biopsies Cut on Frozen Sections) for Biocare Medical.  This kit has long since discontinued by the way...This came from Biocare's Data Sheet.

NOTE:Muscle tissue should be suitably obtained and
frozen within 30 minutes after excision.  The usual
method of freezing is to first mount a transverse section of the muscle on a
chuck using 10% tragacanth gum or equivalent commercial frozen section mounting
media as the adhesive.  The chuck
with the mounted specimen is then immersed in prechilled isopentane until frozen.  Isopentane is precooled when a container of the substance is
placed into liquid nitrogen.  At a
temperature of -160° C, the isopentane has a slightly syrupy
consistency.  Care should be
taken to remove the muscle sample when freezing is complete, usually after 25
to 30 seconds..  Too short a
freezing time produces artifacts; prolonged freezing can produce cracking of
the block.

NOTE: If
the sample cannot be sectioned immediately after freezing, it may be wrapped in
foil and placed in a plastic-lidded container along with a small amount of ice
for moisture.  Specimens may be
stored at -70° C until sectioned.. 
Regards,
Akemi

Akemi Allison-Tacha BS, HT (ASCP) HTL
Histology Manager
Associated Pathology Medical Group Laboratories
105A Cooper Court, Los Gatos, CA 95032 
Cell: (425) 941-4287
E-Mail: akemiat3377 <@t> yahoo..com

--- On Tue, 4/7/09, Bernie Taupin <bernietaupin <@t> ymail.com> wrote:

From: Bernie Taupin <bernietaupin <@t> ymail.com>
Subject: Re: [Histonet] "FREEZY" spray
To: "Jennifer MacDonald" <JMacDonald <@t> mtsac.edu>, "Ingles Claire" <CIngles <@t> uwhealth.org>
Cc: Histonet <@t> lists.utsouthwestern.edu, histonet-bounces <@t> lists.utsouthwestern.edu
Date: Tuesday, April 7, 2009, 9:06 PM

> Does this apply to Liquid Nitrogen as well? I'd hate to have to try to cut fatty sections in the cryostat without it!

I do mostly cryo, and I've never used liquid nitrogen to chill anything within the cryostat. What's that all about? 

I find that it leaves to many ice crystal artifacts anyway. Not cold enough fast enough. You might be making it harder on yourself... just set the temperature of the cryostat for the appropriate temperature of whatever tissue youre cutting, and you should have no need for a crutch, whether it be fluoroethane or liquid nitrogen.

kisses, flowers and rainbows,
Bernie Taupin, King of Cryomicrotomy, Esq.

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