[Histonet] Microtome for autoradiography

[Alan Bright]

Dear Gamal,

You had best look into this a bit deeper. The microtome you are planning
to purchase is not large enough for sectioning whole rats unless they
are only 130mm in length. 
For WBA section you require a cryostat not a bench microtome unless you
would like to place the microtome in a -20ºC cold room.  I am not joking
this is why we started to manufacture this type of cryostat, as Bob
McCulloch at Glaxo, Fulmer Hall, England, 40 years+ ago was doing so, as
he could only stay at this temperature for 1/2 hour max. he asked us to
produce a large sectioning cryostat for him, we did and many 100's were
sold for WBA.

A Tungsten  Carbide Knife will have no problems sectioning through the
undecalcified bone.
We manufacture a range of cryostats for Whole Body Autoradiography

This is an outline of the process for WBA sectioning.

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 4 hours..
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 2 hours.
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 @ 20-30µ depending on
requirements.

I hope you find this useful.

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: abright <@t> brightinstruments.com
Web Site: www.brightinstruments.com


-----Original Message-----
From: gamal akabani hneide [mailto:gakabani <@t> nc.rr.com] 
Sent: 29 September 2003 05:02
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Microtome for autoradiography


Hi All - I need some advice regarding the best microtome to use for
whole
body autoradiography for rats. I am planning to buy a sledge microtome
SM2400 from Leica.
I am going to inject a radiolabeled MAB and sacrifice the animal with
few
minutes to hours, what next? What about the bone sections? Etc.
Any references will be appreciated. Thanks in advanced

Gamal Akabani, Ph.D.
Duke University Medical Center
Department of Radiology
Division of Nuclear Medicine



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