[Histonet] TEM using difficult tissue
Hi I'd like to try some TEM work on rat brain hippocampus. However, the tissue is less than ideal. Does anyone have any information/insight on using old (harvested ~2 years ago) tissue that has been subjected to 20 minutes in sodium sulfide (0.4%), fixed using 30% sucrose in 10% BNF for 48 hours, and then frozen? Thanks very much for any help you can give me. Daphne ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] TEM using difficult tissue
I suspect it will be somewhere between rather ugly to totally worthless, more likely the latter. The only was to know is to find out for yourself. Thaw, rinse multiple times in buffer, osmicate in buffer, rinse, dehydrate, clear and embed in your favorite epoxy. Good luck. Geoff On 9/13/2011 12:17 PM, Daphne Gill wrote: Hi I'd like to try some TEM work on rat brain hippocampus. However, the tissue is less than ideal. Does anyone have any information/insight on using old (harvested ~2 years ago) tissue that has been subjected to 20 minutes in sodium sulfide (0.4%), fixed using 30% sucrose in 10% BNF for 48 hours, and then frozen? Thanks very much for any help you can give me. Daphne ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- -- ** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583 mcaul...@umdnj.edu ** ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] TEM using difficult tissue
Daphne, We do EM work on formalin-fixed, paraffin-embedded tissue occasionally. I will send you a brief protocol under separate cover. Jan Shivers Senior Scientist Histology/IHC/EM Section Head Pathology Teaching Program University of Minnesota Veterinary Diagnostic Laboratory 1333 Gortner Ave. St. Paul, MN 55108 612-624-7297 shive...@umn.edu (Confidentiality Notice: This message, together with any attachments, is intended only for the use of the individual or entity to which it is addressed and may contain confidential or privileged information. If you think you have received this message in error, please advise the sender and then delete this message and any attachments immediately.) - Original Message - From: Daphne Gill dag...@upei.ca To: Histonet@lists.utsouthwestern.edu Sent: Tuesday, September 13, 2011 11:17 AM Subject: [Histonet] TEM using difficult tissue Hi I'd like to try some TEM work on rat brain hippocampus. However, the tissue is less than ideal. Does anyone have any information/insight on using old (harvested ~2 years ago) tissue that has been subjected to 20 minutes in sodium sulfide (0.4%), fixed using 30% sucrose in 10% BNF for 48 hours, and then frozen? Thanks very much for any help you can give me. Daphne ___ 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
