We are trying to establish immunofluorescence techniques using 3D cultures in our laboratory. Does any of your labs are having an optimized method for this?
In our lab, we do that by sectioning the spheroid then treating them as regular tissue, briefly:
- Fix the 3D spheroids with 10% formaldehyde for 2 h,
- Incubate them with 20% w/v sucrose in PBS overnight,
- Wash 3 times with PBS,
- Embed in OCT Cryomount and stored at -80 C,
- Section the spheroids into 5 um-thick slices using a cryostat,
- Store the sections at -80 C until doing the regular IF staining.
Article Quantitative three-dimensional evaluation of immunofluoresce...
Have a look at this article they have an interesting method..
It will vary depending on whether you want to use sections, or do whole-mount for imaging.
- Badges
- Science topic
- Similar topics
- Social Psychology
- Attitude