Has anyone out there analyzed GFP expression from PFA-fixed tissue? (Spleen, liver) How long would GFP stay detectable after fixation? Does it make sense at all to analyze tissues from mice, which are >6 Months old and are sitting in 4% Paraformaldehyde? I would like to see single GFP-positive cells in the tissues and want to analyze where they can be found.
Thanks in advance!
Thomas :-)
Hi Thomas,
In my project, we perfused our rat intracardially, then we harvested the nerve graft and later fixed in 4% PFA for one week (my mistake). followed by embedding in OCT for frozen section. We managed to observed our GFP from each frozen section. No effect on our GFP expression. You may check on this link. Good luck!
https://www.researchgate.net/post/GFP_fluorescence_after_fixation10
Hey Thomas,
I am used to visualize GFP/mCherry in mouse bone marrow after fixation. Normally I fix over night with 1% PFA but also after perfusion and fixation with 2% PFA the signals were super bright. But I never kept the tissue for longer in fixative. After one night in PFA I transfer to sucrose and freeze in OCT. But anyway I would give it a try!
Best Lena
Hi Thomas,
I have to say that I never analyzed mouse tissue kept in PFA for so long.
Usually I follow protocols similar to the one mentioned by other scientists and kept them frozen after cryostat sectioning!
I wouldn't waste time on tissue so old!
Best Paola
- Badges
- Science method
- Similar topics
- Biological Science
- Genetics
- Gene Expression
- Protein Expression