Hello Histonet Users,
I have just started using paraffin and am having many difficulties. Most of the time my tissue crumbles when sectioning. I have no real experience in paraffin histology and have been given the task of becoming proficient by myself, so I am hoping for feedback as to why my tissue keeps crumbling. The tissue in question has been: skeletal muscle, cardiac muscle, liver, and brain (all from rat). The tissue was fixed in 10% neutral buffered formalin for 7 days at 4°C and then transferred to an automated tissue processor, with the following schedule: 2 hours 70% dehydration alcohol 2 hours 80% dehydration alcohol 2 hours 95% dehydration alcohol 2.5 hours 95% dehydration alcohol 2 hours 100% dehydration alcohol 2 hours 100% dehydration alcohol 2 hours 100% dehydration alcohol .5 hour Hemo-De .5 hour Hemo-De .5 hour Hemo-De 1 hour paraffin 4 hours paraffin They were infiltrated for 1 hour without vacuum then embedded. The blocks were stored in the freezer before cutting. The knife angle was 5°. Sections were 5µm thick. I would appreciate any feedback whatsoever. Thank you very much. Michael _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet
