-----Original Message----- From: Chad Miller [mailto:cmil...@labmd.org] Sent: Wednesday, February 03, 2010 4:46 PM To: Delaney, Sondra L Cc: Chad Miller Subject: FFPE Microwave processed biopsies
Microtomy Issue: Our lab has recently switched from using GTF (Glycol Tissue Fixative) as a primary fixative to10%NBF for prostate biopsies. I am experiencing issues with wrinkles, to varying degrees, along the length of the biopsies especially around the lumens of glandular structures. Sections are taken at 3 micrometers with zero visible compression/wrinkling/washboarding etc... of the tissue. When the ribbon is placed on the water bath (47-50 C) the tissue seems to contract and wrinkling appears although, only in the tissue....similar to the coils of a compressed slinky. I have adjusted the water bath temp. increasingly higher until my wax can no longer stand the temp and dissipates around the tissue. This has not cured the wrinkling problem but has shown some improvement. Prostate biopsies that have been fixed in GTF and processed identically to the formalin fixed biopsies are simply perfect in every way. Specimens are embedded using Polyfin (TBS, M.P. 55C) and kept cool for sectioning on ice trays. Sections are collected on charged Histobond slides. Wrinkles appear whether processing solutions are brand new or have been used for previous runs. If anyone has any suggestions ideas or experiences I would sincerely appreciate them. I have included processing times below. Specimen Type: Prostate Needle Core Biopsy Processor: Milestone Histos 5 (Microwave) Specimen Fixative: 10%NBF Specimen Prep: Aligned in linear fashion and sandwiched between sponges to maintain orientation. Max Block Volume: 110 blocks/run Offline Prep: 1. 70% ETOH Rinse (Manual @ 5minutes)--(Changed every 450 blks) 2. 95% ETOH Rinse (Manual @ 5minutes)--(Changed every 450 blks) 3. 100% ETOH Rinse (Manual @ 5minutes)--(Changed every 450 blks) Histos 5 Processor: (0-55blk processing protocol) 1. 100% ETOH (15 minutes @ 65 C)--(Changed every 220 blks) 2. Isopropyl (16 minutes @ 68 C)--(Changed every 220 blks) 3. Vacuum Step (2 minutes ambient) 4. Paraffin (Paraplast M.P.56 C) (23 minutes @ 68 C with Vacuum)-(Changed every 450 blks) This e-mail may contain identifiable health information that is subject to protection under state and federal law. This information is intended to be for the use of the individual named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited and may be punishable by law. If you have received this electronic transmission in error, please notify us immediately by electronic mail (reply). _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
