[Histonet] Help with OCT problem
Has anyone ever experienced OCT blocks that cut like they are soft. I have turned the cyrostat down 10 degrees lower than usual and they are still soft. Really weird part is that I have 20 blocks in this group and some are soft and others are just fine. Tried changing angle, new blade nothing helps. When I finally manage to get a decent section and pick it up on the slide, the OCT around the tissue want even pick up smooth it wrinkles really bad. I tried just a block of OCT that I froze on a chuck and it cut great. I am clueless as to what is going on. Samples were frozen by someone else and brought to me to be cut. Donna Reynolds Core IHC Lab M.D. Anderson Cancer Center, Houston TX Research lab 713-192-8106. Donna Reynolds Core IHC Lab Dept. Cancer Biology, SRB 1.660 713-792-8106 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Help with OCT problem
Donna I have seen soft or rather sticky OCT samples in the past. If you freeze in isopentane and don't let the isopentane evaporate off the samples prior to wrapping the sample in foil, that excess isopentane changes the OCT, it makes it sticky. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Reynolds,Donna M Sent: Thursday, February 10, 2011 12:39 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Help with OCT problem Has anyone ever experienced OCT blocks that cut like they are soft. I have turned the cyrostat down 10 degrees lower than usual and they are still soft. Really weird part is that I have 20 blocks in this group and some are soft and others are just fine. Tried changing angle, new blade nothing helps. When I finally manage to get a decent section and pick it up on the slide, the OCT around the tissue want even pick up smooth it wrinkles really bad. I tried just a block of OCT that I froze on a chuck and it cut great. I am clueless as to what is going on. Samples were frozen by someone else and brought to me to be cut. Donna Reynolds Core IHC Lab M.D. Anderson Cancer Center, Houston TX Research lab 713-192-8106. Donna Reynolds Core IHC Lab Dept. Cancer Biology, SRB 1.660 713-792-8106 ___ 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] Help with OCT problem
Donna, I would also check to make sure that the tissue wasn't held in an something with a lot of alcohol, like RNA later. I found out first hand that it doesn't work well. Good luck On Thu, Feb 10, 2011 at 11:38 AM, Reynolds,Donna M dreyn...@mdanderson.org wrote: Has anyone ever experienced OCT blocks that cut like they are soft. I have turned the cyrostat down 10 degrees lower than usual and they are still soft. Really weird part is that I have 20 blocks in this group and some are soft and others are just fine. Tried changing angle, new blade nothing helps. When I finally manage to get a decent section and pick it up on the slide, the OCT around the tissue want even pick up smooth it wrinkles really bad. I tried just a block of OCT that I froze on a chuck and it cut great. I am clueless as to what is going on. Samples were frozen by someone else and brought to me to be cut. Donna Reynolds Core IHC Lab M.D. Anderson Cancer Center, Houston TX Research lab 713-192-8106. Donna Reynolds Core IHC Lab Dept. Cancer Biology, SRB 1.660 713-792-8106 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Patrick Laurie HT(ASCP)QIHC CellNetix Pathology Laboratories 1124 Columbia Street, Suite 200 Seattle, WA 98104 plau...@cellnetix.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Help with OCT problem
I agree with previous responders to this query, as I've gotten alcohol on my blocks and since alcohol doesn't freeze real well it makes a sticky mess with the OCT. The affected OCT can be scraped off with a razor blade. Hopefully the tissue itself did not come in contact with alcohol prior to embedding. Regards, Merced --On Thursday, February 10, 2011 12:51 PM -0800 Patrick Laurie foreig...@gmail.com wrote: Donna, I would also check to make sure that the tissue wasn't held in an something with a lot of alcohol, like RNA later. I found out first hand that it doesn't work well. Good luck On Thu, Feb 10, 2011 at 11:38 AM, Reynolds,Donna M dreyn...@mdanderson.org wrote: Has anyone ever experienced OCT blocks that cut like they are soft. I have turned the cyrostat down 10 degrees lower than usual and they are still soft. Really weird part is that I have 20 blocks in this group and some are soft and others are just fine. Tried changing angle, new blade nothing helps. When I finally manage to get a decent section and pick it up on the slide, the OCT around the tissue want even pick up smooth it wrinkles really bad. I tried just a block of OCT that I froze on a chuck and it cut great. I am clueless as to what is going on. Samples were frozen by someone else and brought to me to be cut. Donna Reynolds Core IHC Lab M.D. Anderson Cancer Center, Houston TX Research lab 713-192-8106. Donna Reynolds Core IHC Lab Dept. Cancer Biology, SRB 1.660 713-792-8106 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Patrick Laurie HT(ASCP)QIHC CellNetix Pathology Laboratories 1124 Columbia Street, Suite 200 Seattle, WA 98104 plau...@cellnetix.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician III Cardiovascular Medicine 348 Biomedical Research Building State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 USA lei...@buffalo.edu 716-829-6118 (Ph) 716-829-2665 (Fx) No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Help with OCT problem
Same with ETOH. We wipe our surfaces down with it to clean them so no cross contamination as we freeze our skin here on the metal side areas of our cryostats. If the alcohol hasn't been wiped dry or evaporated we have that problem too. Claire From: histonet-boun...@lists.utsouthwestern.edu on behalf of Liz Chlipala Sent: Thu 2/10/2011 1:49 PM To: Reynolds,Donna M; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Help with OCT problem Donna I have seen soft or rather sticky OCT samples in the past. If you freeze in isopentane and don't let the isopentane evaporate off the samples prior to wrapping the sample in foil, that excess isopentane changes the OCT, it makes it sticky. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
