Hi,
does anyone know why TRAP staining faids in FFPE rat femur sections?
My protocol: After deparaffination and rehydration i put the slides in Acetat Buffer for 20min, then they go to the prewarmed staining solution (Napthol AS-MX Phosphate (Sigma N-4875) and Fast Red TR hemizinc salt (Sigma 368881) in a 37° oven for 1h. Then i wash in aqua dest and counterstain with Meyers hematoxylin, aqua dest wash, ~10 dips in ammoniumwater, aqua dest wash and mount them with aquatex.
The stain intensity was great but the next day the intensity is much lower and it keeps faiding... It the annoying thing is that I already established this staining, which worked fine and now with my good slides this happend....
Thanks in advance for your tips!
As far as I remember this staining is water-soluble so using xylene-based mounting media instead of water-based aquatex may be a good idea.
Hi Anton Lennikov . Thank you for your answer! That would make a lot of sense and I will try to change the mounting media to a xylol bases one!
For long-term storage, you may also want to dehydrate your slides after staining using grading ethanol->xylene before mounting with xylene mounting media.
Yes i usually alwas do that with xylen based mounting media. We also scan our slides so we have the whole tissue as 40X digitally stored.
- Badges
- Science method
- Similar topics
- Biological Science
- Histology
- Staining