Re: [Histonet] FW: cutting a 5um frozen section from a 50um section of brain
I just made the aluminum temple from a 1/2 inch 6x6 piece on aluminum from Home Depot. With the sheet you will este,Italy make it look like a clock face using a sharpe. Why you want to do this is i found the aluminum plates have enough variance that if I did not always use the same side it changed the plain of my cut. The key is to make sure once you established correctly you can repeat everything exactly the same. Sent from my iPad On Oct 6, 2017, at 4:44 PM, Dessasau III, Evan mailto:edde...@emory.edu>> wrote: 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 mailto:edde...@emory.edu>> Cc: Siravo, Michele L. mailto:michele.sir...@thermofisher.com>>; Cole, Alan D. mailto: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<mailto: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
Re: [Histonet] FW: cutting a 5um frozen section from a 50um section of brain
Hi Loralei, I will see If I can find someone with a sliding microtome. Are you using OCT to block the 50 um section? How do you ensure the section stays flat while blocking/freezing? Thank you for your reply ! E-van From: Loralei Dewe [mailto:lld...@gmail.com] Sent: Friday, October 06, 2017 6:13 PM To: Dessasau III, Evan Subject: Re: [Histonet] FW: cutting a 5um frozen section from a 50um section of brain I would use a sliding microtome and freeze the 50 um sections solid with dry ice. I used to do sections of mouse brain that way. Loralei On Oct 6, 2017 9:10 AM, "Dessasau III, Evan via Histonet" mailto:histonet@lists.utsouthwestern.edu>> wrote: 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
Re: [Histonet] FW: cutting a 5um frozen section from a 50um section of brain
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 Cc: Siravo, Michele L. ; Cole, Alan D. 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
Re: [Histonet] FW: cutting a 5um frozen section from a 50um section of brain
HI Ms. Powell!! , Thanks for the reply. When we last all saw each other at the Georgia Histopalooza I expressed an interest in training on the cryostat with Cindy Baranowski. She was so nice and had me come over to the hospital back in June. That was really really nice of her! But now I can't get the new Cryostat smell out of my nose! I have not reached out to her nor Jennifer nor Jeanine with this issue. I bother Jennifer all the time I try not to tread too much, she too is very helpful. There is another tech Susan Jenkins here at Yerkes that I have talk to about my project. She was doing something similar but found using the Vibratome worked for her project. I need sections thinner than the vibratome will cut. I'm using the Leica CM3050 S. I'm able to get the temp right to cut brain sections with little curl to them. I'm talking about getting my 50 um section(they are about 2 to 3cmsq) to remain flat through freezing so that I cut a section in one plane. I hope that makes a little more sense. I will reach out to Cindy. Thank you so much! E-van -Original Message- From: Shirley A. Powell [mailto:powell...@mercer.edu] Sent: Friday, October 06, 2017 1:41 PM To: Dessasau III, Evan Subject: RE: [Histonet] FW: cutting a 5um frozen section from a 50um section of brain Hi Even, Have you asked any of the other histotechs at Emory about this. I know Cindy Baranowski, Jeanine Bartlett and Jennifer Smith work there. What cryostat are you using. I know that the temp for cutting brains is a little different from other tissues. Sometimes the machines have anti-roll systems in place too. Also How big are your brain sections? Shirley Powell -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 http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] FW: cutting a 5um frozen section from a 50um section of brain
Haley, Maybe ask Stephen directly. He is on histonet, and has a website: https://www.pathologyinnovations.com/ Tim Morken Pathology Site Manager, Parnassus Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of Pathology UC San Francisco Medical Center -Original Message- From: Haley Huggins via Histonet [mailto:histonet@lists.utsouthwestern.edu] Sent: Friday, October 06, 2017 10:31 AM To: Dessasau III, Evan Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] FW: cutting a 5um frozen section from a 50um section of brain I would also be interested in knowing this tip if anyone knows it. I will also have to check out that book. We only do cryostat sections at our lab. *Haley Huggins, HT (ASCP)cm* *Technical Lab Supervisor* *1050 Las Tablas Rd, Suite 14* *Templeton, CA 93465* *Office: 877-230-1518* On Fri, Oct 6, 2017 at 9:09 AM, Dessasau III, Evan via Histonet < histonet@lists.utsouthwestern.edu> wrote: > 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 > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ___ Histonet mailing list 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
Re: [Histonet] FW: cutting a 5um frozen section from a 50um section of brain
I would also be interested in knowing this tip if anyone knows it. I will also have to check out that book. We only do cryostat sections at our lab. *Haley Huggins, HT (ASCP)cm* *Technical Lab Supervisor* *1050 Las Tablas Rd, Suite 14* *Templeton, CA 93465* *Office: 877-230-1518* On Fri, Oct 6, 2017 at 9:09 AM, Dessasau III, Evan via Histonet < histonet@lists.utsouthwestern.edu> wrote: > 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 > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
