I'm having a lot of trouble sectioning nerve tissue (sciatic and ovarian). I can get any decent longitudinal cuts, the tissue catches and shreds or completely disintegrates in the water bath. I can get some cross sections, but once they are mounted on the slide, I find that they are separating and melting out from the surrounding paraffin. I am beyond frustrated at this point. Any help would be greatly appreciated.
Have you checked the water bath and the slide warmer temperature?
I sometimes have the same issue with mature gonads when the water is too warm.
My water bath is at 38C - when I can actually get a section, I've used lower temperatures (down to 30C) to try and figure out the optimal temperature, but as soon as I do that, the tissue no longer adheres when staining.
As for the fixation, the tissue was fixed in the 10% formalin for 48 hours. I've found that with this tissue 24 is too short. I'm not sure what the lab I'm working with has available. How long would you recommend fixation in the PFA?
Thank you for your help on this Carlos and Markus!
Thanks Markus, that's really helpful! - I suppose I should mention these are human cadaver tissue. That might be relevant
- Badges
- Science topic
- Similar topics
- Correction