RE: Whole mouse snap freezing for frozen sections2

From:Alan Bright






Dear Gayle,

We manufacture a range of instruments for whole body sectioning, and in the
main this is the method used for whole mouse and rat snap freezing for
frozen sections.

The appropriate safety equipment must be used e.g. eye protection and
gloves.

1) Pin the specimen by the limbs onto a flat 3mm or 1/8" sheet of hardboard.
2) Immerse the above into a slurry of Petroleum Ether and Solid C02
solution, for at least 30 minutes.
3) Remove from hardboard and remove limbs and tail.
4) Wash fur with a 0.07% solution of Trinton X.
5) Immerse specimen into a mould and fill with a 2% solution of Carboxy
Methyl Cellulose.
6) Finally immerse the above into a slurry of Petroleum Ether and Solid C02
solution, for at least 30  minutes.
7) Remove from mould and mount the moulded specimen onto a object holder for
mounting onto the microtome using a 2% solution of Carboxy Methyl Cellulose.
8) Using a Tungsten tipped knife trim specimen until the required level is
reached, adhere 3m's or equivalent section mounting tape to the face of the
sample and cut the required section @ 20-30 depending on requirements.

This can be achieved for mice using a Bright OTF5000 fitted with motorized
drive and a large area sectioning kit at a fraction of the cost of the type
of cryostat you mention in your email of today on Histonet.There is no need
for an expensive tape transfer system for the retrieval of sections as the
standard 3m's or equivalent section mounting tape has been well proven over
the past 35 years.
For larger specimens we manufacture another two cryostats 8250 and 9400. A
brief description can be viewed on our website brightinstruments.com

Best Regards

Alan Bright

Bright Instrument Co.Ltd.
St Margaret's Way
Huntingdon
Cambridgeshire
PE29 6EU
England

Tel No:+44 (0)1480 454528
Fax No:+44 (0)1480 456031
Email: AlanBright@brightinstruments.com
Web Site: brightinstruments.com





-----Original Message-----
From: Gayle Callis
Sent: 25 June 2002 21:38
To: histonet@pathology.swmed.edu
Subject: Whole mouse snap freezing for frozen sections


I was asked how people snap freeze a euthanized whole mouse for serial
frozen sections.  I've snap frozen whole mouse heads, tibias, femurs using
dry ice cooled hexanes after coating the samples with either diluted OCT or
4% PVA, 30,000 - 70,000 water soluble from Sigma. Is this snap freezing
method going to be too slow for a whole mouse. Any help would be
appreciated.  Off to PubMed.

Searching the Archives was not turning up the answer




Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
19th and Lincoln St
Bozeman MT 59717-3610

406 994-6367 (voice mail)
406 994-4303 (FAX)

email: UVSGC@montana.edu






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