The fixation time is too short. Fix as long as you can (how about 2-3 days at least for brain?)
Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Poole, Kimberly Sent: Wednesday, 28 July 2010 12:28 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Brain Problems Hi there, I am currently using rat brain for histology but something is not working. I follow the protocol below for tissue processing. Station Solution Routine (mins) 1 Formalin 10 2 Formalin 10 3 70% Ethanol 60 4 80% Ethanol 240 5 96% Ethanol 240 6 100% Ethanol 240 7 Xylene 120 8 Xylene 120 9 Xylene 120 10 Paraffin Wax 120 11 Paraffin Wax 180 12 Paraffin Wax 180 TOTAL TIME 27 hours 20 minutes I embedded my brains and then cut them afterwards. When the brain section hits the water, it looks like a blob of fat on the water. Then it spreads reall, really slowly. If a cut is made to the paraffin the brain bursts open and it falls apart. Am I doing something wrong in the processing stage? Is the tissue dehydrated too much. Any feedback would be greatly appreciated. Kimberly Poole B.Sc Casualty Management Section | Section de la gestion des blessés Defence Research and Development Canada Suffield | Recherche et développement pour la défense Canada Suffield Medicine Hat, AB, Canada T1A 8K6 kimberly.po...@drdc-rddc.gc.ca <mailto:kimberly.po...@drdc-rddc.gc.ca> Telephone | Téléphone 403-544-5347 / Facsimile | Télécopieur 403-544-4714 Government of Canada | Gouvernement du Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************************* This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************************* _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet