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, E-van From: Jacox, Robert A. [mailto:robert.ja...@thermofisher.com] Sent: Friday, October 06, 2017 12:55 PM To: Dessasau III, Evan <edde...@emory.edu> Cc: Siravo, Michele L. <michele.sir...@thermofisher.com>; Cole, Alan D. <alan.c...@thermofisher.com> Subject: RE: [Histonet] FW: cutting a 5um frozen section from a 50um section of brain
Evan, 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 Robert Jacox Commercial Marketing Manager Anatomic Pathology Thermo Fisher Scientific Tel: 269-544-5651 l Mobile: 269-598-0747 robert.ja...@thermofisher.com<mailto:robert.ja...@thermofisher.com> l www.thermoscientific.com<http://www.thermoscientific.com> -----Original Message----- From: Dessasau III, Evan via Histonet [mailto:histonet@lists.utsouthwestern.edu] Sent: Friday, October 06, 2017 12:09 PM To: 'histonet@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. Thank you, E-van 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)727-7744 lab (404) 727-7902 office ________________________________ This e-mail message (including any attachments) is for the sole use of the intended recipient(s) and may contain confidential and privileged information. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message (including any attachments) is strictly prohibited. If you have received this message in error, please contact the sender by reply e-mail message and destroy all copies of the original message (including attachments). _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu<mailto:Histonet@lists.utsouthwestern.edu> http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
