I am learning how to do negative staining on cells for TEM, and I am seeing highly variable shapes/arrangement of nuclei. Is this normal, or did something go wrong with the processing?
You are performing positive (not negative) staining.
Nuclei are not spherical and can have varied shape.
I agree with Vladimir (positive staining) is indicated in the images you are displaying. Nuclei can be near circular in sectional view, but the more common shape is non-round. Many can be highly convoluted, almost Amebia-like. The same shapes can be detected with SEM (with the right protocol), but more importantly, cryo-sectioning of cryo-fixed tissue produces the same convoluted shapes. Cryo-TEM was, for many years, considered the go-to method for proofing chemical fixation methods. But, since we have perfected chemical fixation methods, cryo-techniques are now much less commonly done.
If your fixative pH is correct for the tissue, and your osmolarity is correct, you should get good chemical fixation results.
Yes. I agree with Vladimir Dusevich.
- Badges
- Science topic
- Similar topics
- Psychology
- Mental Processes
- Learning