[Histonet] -80 freezer question
JWatson <@t> gnf.org
Wed Nov 20 10:18:26 CST 2013
I do not remember exactly what study or publication I got this from, but I had to do some research a few years ago because our lab ops department did not want to have our freezer set at -80. I came up with a few reasons:
. Some proteins have been shown to break down at -60 but be stable at -80
. DNA and mRNA recovery is better at -80 vs. -60
. Some antibodies are better preserved at -80
o Our Collagen X antibody from Abcam (ab58632) requires -80 storage
Diagn Mol Pathol. 1992 Mar;1(1):73-9.
Role of the frozen tissue bank in molecular pathology.
Naber SP, Smith LL Jr, Wolfe HJ.
Department of Pathology, Tufts University School of Medicine, New England Medical Center Hospitals, Boston, MA 02111.
The new discipline of molecular pathology requires that high-quality, intact genomic DNA, mRNA, and proteins be available from frozen tissue samples. It is now necessary for pathology laboratories to establish consistent guidelines for the preparation and storage of frozen tissue samples in order to have properly preserved tissues available for diagnostic molecular techniques. Maintaining a frozen tissue bank requires a pathologist to oversee this program and to integrate it into the routine surgical pathology activities. A member of the laboratory technical staff can serve as a tissue bank coordinator and have responsibility for preparation of tissue samples, their systematic storage and retrieval, and routine maintenance of equipment and supplies. Tissue sampling must be done as soon as possible after excision of the specimen and is the responsibility of a qualified pathologist. The samples may be snap frozen without cryoprotection at -78 degrees C or colder for subsequent use in procedures requiring the extraction of genomic DNA, mRNA, or protein. To preserve tissue architecture and cytologic features for immunohistochemistry and in situ hybridization, the tissue should be frozen at -78 degrees C or colder with a cryoprotectant such as OCT. Long-term storage of the frozen tissue is recommended at -140 degrees C or colder in a locked liquid nitrogen freezer, and the record of sample inventory can easily be kept in a computerized database. Tissues sampled and stored under these conditions have been used successfully in a wide variety of molecular techniques. In addition to malignant tumor tissue, samples from benign lesions and normal tissues should be frozen.(ABSTRACT TRUNCATED AT 250 WORDS)
[PubMed - indexed for MEDLINE]
James Watson HT ASCP
GNF Genomics Institute of the Novartis Research Foundation
jwatson <@t> gnf.org
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Paula Sicurello
Sent: Tuesday, November 19, 2013 2:23 PM
Subject: [Histonet] -80 freezer question
What is the rationale for storing frozen patient samples at -80? Is there a range that keeps the sample viable but not as cold?
Our -80 (which we just inherited) is struggling and I would like to raise the temperature a smidge to ease the stress on the compressor.
Paula Sicurello, HTL (ASCP)
Supervisor, Clinical Electron Microscopy Laboratory Duke University Health System Rm.#251M, Duke South, Green Zone Durham, North Carolina 27710
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