[Histonet] -80 degrees rationale

Pam Marcum mucram11 <@t> comcast.net
Wed Nov 20 10:07:55 CST 2013

Also we had time when finding a non-defrost freezer was difficult and lower temps were not only best; they were about the only way not to have unit turn off and on allowing tissue and antibodies to partially defrost.   Some of us learned that the hard way in early days of IHC. 


Bill is correct in his answer about why it should be stored at the various temps also. 


Pam Marcum 


----- Original Message -----
From: "WILLIAM DESALVO" <wdesalvo.cac <@t> outlook.com> 
To: "Paula Sicurello" <patpxs <@t> gmail.com>, "histonet" <histonet <@t> lists.utsouthwestern.edu>, "Microscopy <@t> microscopy.com" <microscopy <@t> microscopy.com> 
Sent: Wednesday, November 20, 2013 9:58:42 AM 
Subject: RE: [Histonet] -80 degrees rationale 

Here is my stab at why -80 C. 
Temperatures between 0°C and −25°C, the enzymatic activity of cells is only slowed but remains active. Below −40°C physiochemical exchanges are frozen. Cellular morphology is preserved at -80°C. Shelf life of tissue increases as the temperature drops. Once you get below -80°C you will need cryoprotectors and when you use cryoprotectors, temperatures must be below −130°C and will go as low as −196°C (liquid nitrogen).  Antibodies and proteins in solution are stable at −20°C. 

William DeSalvo, BS HTL(ASCP) 

> Date: Wed, 20 Nov 2013 09:25:54 -0500 
> From: patpxs <@t> gmail.com 
> To: histonet <@t> lists.utsouthwestern.edu; Microscopy <@t> microscopy.com 
> CC: 
> Subject: [Histonet] -80 degrees rationale 
> Hello My Fellow Listers, 
> The question of the day is:  What is the rationale for storing frozen 
> biopsies at -80 degrees? 
> I have seen protocols that range in temperature from -40 to -80 degrees. 
> Was -80 selected because that was the lowest freezers could go back in the 
> day? 
> Awaiting your chilly responses! 
> Thanks in advance, 
> Paula 
> -- 
> Paula Sicurello, HTL (ASCP) 
> Supervisor, Clinical Electron Microscopy Laboratory 
> Duke University Health System 
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> Durham, North Carolina 27710 
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