Hello there,
The literature is plenty of examples of immunostaining in live cells, but is hard to find the same in live organotypic culture. Does anyone have experience with this? Is there (a priori) any caveat about this technique?
Thanks,
J
Organoids should be fixed with 10% formalin for 24-48 hours and then sliced for the immunostaining. Some researchers including me perform the whole-mount staining, but this method requires the relatively a high amount of antibody. By contrast, the suggested method does not need a high amount of antibody.
If you want to stain live cells, formalin fixation is not possible. After 2 days in formalin everything will be dead.
I could imagine that it depends on the cellular position of the antigen you want to stain: Staining cell surface proteins will be much more likely than proteins sitting in the mitochondrial membrane for example as you need to get access to the epitope.
Good luck, it sounds like an interesting but difficult idea!
Hello
I agree with researchers and the article here will help you
Article Maintaining Live Cells and Tissue Slices in the Imaging Setup
Good Luck
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