Dear members, first I wanna thank all the people who responded to my first email (Andrea, Mesru and Emily).Your tips were very helpful. Unfortunately I am still fighting agains bubbles/ wrinkles/bubble-shaped wrinkles between my slices and the slide.
During the last couple of days I tried: i) to adjust the temperature of the chamber and the sample, seems like -16°C brings the best slices ii) to cool the slide by putting it into the chamber and warming it with my thumb just the second I held it towards the slice to adher iii) to warm the slides by putting them on a 37°C warm heating-plate after attachement iv) to moisten the slides with a little bit of bidest before letting the slices attach and put these slides on a 37°C heating plate afterwards...that was kind of desperate try to straighten the slices slowely v) 3 different intermedia before embedding the sample in pure Tissue Tek, a) 30% Succrose, b) half 30% succrose and half Tissue Tek, c) half Tissue Tek half PBS but...nothing really helped. I still have wrinkles aka bubbles. Tomorrow I am going the cut perfusion fixed rat spinal cord, hoping that this makes a difference. Anyway, I think I have some options left 1) trying to cut on a different mikrotome and 2) Leica promised to send me some slide/embedding solution/blade samples that worked for other groups. Can anybody think of something I havent tried yet? Besides of raising slide thickness and cutting for freefloating procedure? I would be so thankful for any ideas/suggestions/tips! Regards Christina _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet