Is there a maximum section thickness recommended for slide-mounted immunofluorescence protocols? Our experiments involve 30um mouse brain sections fixed in 4%PFA and sectioned on the freezing microtome.
you may e-mail to this expert, they do regularly:
Del Negro, Christopher A
If you look up Amisha Parmar from University of Central Lancashire on Research Gate, she asked a similar question. The answers provided are very insightful and I recommend taking a look. Hope this helps!
From all the work I have done paraformaldehyde would cause auto-fluorescence. With sections that are 30um, I would recommend doing a floating staining procedure in small glass staining dishes or petri dishes. moving the sections from one dish to another with a slide, letting the section float in each substrate and buffer wash. Most crysections I've worked with were fixed in acetone or alcohol to retain morphology of the cells.
- Badges
- Science topic
- Similar topics
- Biological Science
- Histology
- Sectioning