Hi there! I work in a pathology lab where we section paraffin-embedded drosophila heads at 5um. My goal was to do it at 2um, but it was just impossible. After working for 4 months, I decided to section at 5um. But it’s still so difficult! The major problem I see right now is that the tissue isn’t adhering to the slide. I use charged slides, I keep my water at 38C. Once I scoop up the ribbons onto my slide, I let it set on the heating block to dry and then keep it in the incubator at 37C overnight. I do an H&E where I deparaffinize the slides in xylene, keeping them in two separate containers for 10 minutes each. I then rehydrate them (100%, 95%, and 70% ethanol, then water), keep them in hematoxylin, a quick rinse in di acid, and eosin, and then dehydrate them. Here are the problems I’m facing to be specific- The tissue seems to be attached well post-slide mounting and after drying overnight. But there’s a lot of the tissue sliding off, migrating, or folded and shredded when I check it after staining.

My goal is that I need to have cross-sections with intact exoskeleton and brain (especially) that are publication-worthy.

I’ve never sectioned at such a low thickness and I’ve been struggling for 6 months. My PI and I are stressed, please let me know if you have any pointers that can help, thanks!