[Histonet] FW: cutting a 5um frozen section from a 50um section of brain
Dessasau III, Evan
eddessa at emory.edu
Fri Oct 6 15:44:45 CDT 2017
Hi Robert, thank you for your reply. So you froze the section on the aluminum with the "clock face" and then put this inverted, in the Microm 560 mold? ours is an older unit that has been around and not all the parts came with it so I don't have the 90 degree block chuck. Is this the fixed head you speak of?
I love the idea of a control block for cutting I image you mean. I have been putting a layer of OCT on top of the block/section after I pop it out of the mold. I use this "extra" OCT to orientate my x-y.
Robert how thick was the section you were trying to cut?
I'm wondering if anyone knows if samples move about as the OCT freezes?
Thank you again,
From: Jacox, Robert A. [mailto:robert.jacox at thermofisher.com]
Sent: Friday, October 06, 2017 12:55 PM
To: Dessasau III, Evan <eddessa at emory.edu>
Cc: Siravo, Michele L. <michele.siravo at thermofisher.com>; Cole, Alan D. <alan.cole at thermofisher.com>
Subject: RE: [Histonet] FW: cutting a 5um frozen section from a 50um section of brain
I had a similar project and did after a lot of trial and error was able to get consistent results. The way I approached it was the following. There are several variables that need to be accounted for.
1. Is the tissue cut on the plain I want to see (this has to be yes)
2. Mounting the section to a known flat surface
3. Attaching the section to the chuck in a consistent manner
4. The cutting angle of the specimen head
5. The placement of the knife
6. The accurate insertion of the knife/blade
For point 2 I used a flat piece of aluminum and drew clock face marks on it. This way I knew I was consistently attaching the section on a known plain. Point 3 I used some embedding molds from the Microm 560 to act as a known way to attach the specimen. The molds end up making a bullet shaped sample so it was easy to keep orientation (full disclosure I would for Thermo (Microm) I would guess any consistent type mold will do). Point 4 I went from an adjustable head to a fixed head on the cryostat. Points 5 and 6 - I created a control block that I cut prior to beginning to cut and any time I moved the knife holder or changed the blade. For this I saved old 100 micron sections and would embed one using the Point 1-4 methodology and verify the plain.
I hope this helps and let me know if you need further assistance
Commercial Marketing Manager
Thermo Fisher Scientific
Tel: 269-544-5651 l Mobile: 269-598-0747
robert.jacox at thermofisher.com<mailto:robert.jacox at thermofisher.com> l www.thermoscientific.com<http://www.thermoscientific.com>
From: Dessasau III, Evan via Histonet [mailto:histonet at lists.utsouthwestern.edu]
Sent: Friday, October 06, 2017 12:09 PM
To: 'histonet at lists.utsouthwestern.edu'
Subject: [Histonet] FW: cutting a 5um frozen section from a 50um section of brain
Hi Histonet , I have been trying to cut a FLAT 4 to 10 um frozen section from 50um sections of brain. Every time I think I have the tissue flat the sections are never in the same plane. I found a wonderful book in pdf format(A Practical Guide to Frozen Section Techniques, Stephen R. Peters) online with lots of wonderful tips but I'm having no luck implementing the tips. Has anyone ever tried this? Any help is GREATLY appreciated.
E-van D. Dessasau, III, HTL(ASCP)cm
Supervisor, Histology Division of Pathology Emory University Yerkes NPRC Main Center Rm. 2122
954 Gatewood Rd.
Atlanta, GA. 30329
(404) 727-7902 office
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