Hi Dorianne, You need to freeze your tissue faster. Ideally, isopentane placed in a metal cup, that is that is then frozen in a dewar of liquid nitrogen, works best. The isopentane, once frozen, is thawed a little with a metal rod to produce a small liquid pool and your tissue is placed in this for about one minute. You need some equipment for this procedure, such as the metal cup that can sit inside a small, open dewar of liquid nitrogen. Alternatively, you can freeze directly in liquid nitrogen, though you need to beware of the tissue fracturing due to the sudden and extreme temperature reduction. Slower freezing of tissue (sitting on dry ice, etc.) allows ice crystals to form in the tissue, creating the vacuoles you describe. I hope this helps. Phil.
Philip Manfre, BA, HT (ASCP) Associate Principal Scientist Merck Research Laboratories WP81-406 770 Sumneytown Pike West Point, PA 19486 215-652-9750 philip_man...@merck.com -----Original Message----- From: Bonello Dorianne M at Health-MDH via Histonet <histonet@lists.utsouthwestern.edu> Sent: Friday, July 16, 2021 11:25 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] frozen section problem EXTERNAL EMAIL – Use caution with any links or file attachments. Dear all, We are experiencing freezing artifacts on our frozen sections. Basically, we are seeing cavity-like structures under the microscope, mostly elongated, especially when it's a frozen section on brain tissue. This is most probably happening due to ice crystal formation. We're not using cryospray, relying only on the cryobar boost function. Does anyone has a solution to this problem please? Regards, Dorianne Bonello Allied Health Practitioner (MLS) Histology Laboratory - Pathology Health-Mater Dei Hospital [cid:image001.jpg@01D67184.63288530] T +356 +356 25456434 E dorianne.m.bone...@gov.mt Mater Dei Hospital, Triq id-Donaturi tad-Demm, l-Imsida, Malta MSD 2090 | Tel +356 2545 0000 | https://deputyprimeminister.gov.mt/en/MDH/Pages/Home.aspx<https://deputyprimeminister.gov.mt/en/MDH/Pages/Home.aspx> | https://www.facebook.com/materdeihospital/ Think before you print. Kindly consider your environmental responsibility. This email and any files transmitted with it are confidential, may be legally privileged and intended solely for the use of the individual or entity to whom they are addressed. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (2000 Galloping Hill Road, Kenilworth, New Jersey, USA 07033), and/or its affiliates Direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet