[Histonet] "FREEZY" spray

Wed Apr 8 03:05:39 CDT 2009

Say thank you Uncle Kemlo!!

Kemlo Rogerson	
e-mail kemlorogerson <@t> nhs.net if not at work.			
DD   01934 647057 or extension 3311     Mob 07749 754194; 
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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Tony Henwood
Sent: 08 April 2009 08:58
To: Bernie Taupin; Akemi Allison-Tacha; Jennifer MacDonald; Ingles Claire
Cc: Histonet <@t> lists.utsouthwestern.edu; histonet-bounces <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] "FREEZY" spray

Hey,

Would you believe that putting the word "Taupin" into my outlook rules actually works.
Now every obnoxious post from "Taupin" actually disappears into my Junk folder and I do not have to read the drivel.

Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) 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 Bernie Taupin
Sent: Wednesday, 8 April 2009 3:38 PM
To: Akemi Allison-Tacha; Jennifer MacDonald; Ingles Claire
Cc: Histonet <@t> lists.utsouthwestern.edu; histonet-bounces <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] "FREEZY" spray

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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