[Histonet] RE: Inconsistent Sections with Cryostat

[Manfre, Philip]

I believe you may need to have the unit serviced.  It sounds like something is not tight enough, perhaps the stage or blade holder unit.  You said you secured everything which makes me think you have some issue with the cryostat itself.  If a sharp blade, tightened blade and specimen, and varying the speed doesn't work, then you may need professional assistance form a service technician, especially if it is an older unit that has been moved around.  Have you tried different temperatures?  I don't think that is the problem but it is worth a try. 

Good luck! Cryosectioning can be an art in itself. Those decent sections are likely thicker than you intend, by the way.  Classic "thick and thin".


Philip Manfre, B.A., HT (ASCP)
Associate Principal Scientist
Merck Research Laboratories
WP45-251
PO Box 4
West Point, PA 19486

215-652-9750
215-993-0383 (fax)
philip_manfre <@t> merck.com




-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Perow, Elliott S (PEROWES10)
Sent: Thursday, October 03, 2013 12:33 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Inconsistent Sections with Cryostat 

Hello All,

I have a Leica Cryocut 1800 with a Reichert-Jung 2020 microtome that has sat for 8 years and went through a move from one academic building to another.  I am trying to section brook trout brain, but am having difficulty getting consistent sections.
I will get a good section and then the next section will be a very very thin shaving of usually just the top portion of the OCT block.  I have tried adjusting the clearance angle, made sure the blade and specimen is secured, replaced the blade with a new one, have tried different sectioning speeds, different thicknesses of the section and still continually only get a decent section every other turn.  I was wondering if anyone has had any similar experiences or if maybe the machine just needs a maintenance check due to it being an older machine.  I wasn't sure if the advancement mechanism may be off or if it is another issue that doesn't seem apparent to me.  Any help would be greatly appreciated.

Thanks,

Elliott Perow
Juniata College
Biology POE


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