Dear Histonet Colleagues, I am trying to develop a processing protocol for our Leica TP1020, but the first run has not been very successful. The specimen is canine femur (section lengths of 1" to 1-1/2 "). Following is a list of the processing reagents and times for my first attempt: 70% ETOH for 4 hours with vacuum 95% ETOH for 4 hours with vacuum 95% ETOH for 4 hours with vacuum 95% ETOH for 4 hours with vacuum 100% ETOH for 8 hours with vacuum 100% ETOH for 8 hours with vacuum 100% ETOH for 8 hours with vacuum 100% ETOH for 8 hours with vacuum Methyl Methacrylate (Pure) for 4 hours with vacuum Methyl Methacrylate (Pure) for 6 hours with vacuum Methyl Methacrylate (Pure) for 8 hours with vacuum Methyl Methacrylate with Dibutyl Phthalate and Perkadox for 4 hours with vacuum Methyl Methacrylate with Dibutyl Phthalate and Perkadox for 4 hours with vacuum (specimen was placed into refrigerator overnight) Methyl Methacrylate with Dibutyl Phthalate and Perkadox for 4 hours with vacuum Methyl Methacrylate with Dibutyl Phthalate and Perkadox for 4 hours with vacuum (specimen was placed into refrigerator overnight)
Methyl Methacrylate with Dibutyl Phthalate and Perkadox for 4 hours with vacuum Methyl Methacrylate with Dibutyl Phthalate and Perkadox for 4 hours with vacuum (specimen was placed into refrigerator overnight) Specimen was removed from refrigerator and embedded in a water bath for 2 days. I cut the specimen in half in the longitudinal direction. The cortices appear to have been infiltrated fairly well, but the marrow still has some very large holes. If anyone would happen to have a protocol that works well with this type of tissue and this processor, it would be greatly appreciated if you could offer your advice and expertise as to what I should do next. I do have one more specimen still running in the processor and will probably embed it on Monday. The first specimen was started on a Monday and embedded the following Monday (approx. 8 days), so this second specimen will have been in MMA for an extra 8 days before embedding. Thank you very much for your help. Jim _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet