We are trying to stain cholangiocytes with mitotracker green and perform confocal live imaging. However, once we added mitotracker, the entire monolayer of cells started to peal off from the bottom of the well. We use Ibidi coated 8-well plates. We tried different concentrations of mitotracker, using collagen to coat the well, but nothing had worked. The same thing happened with mitoSOX. JC-1 dye works fine. Any suggestion is highly appreciated!
Hi Chunyue,
I don't have hands-on experience in culturing cholangiocytes, but I used to coat my plates with fibronectin or Matrigel for endothelial progenitor cell culture. From my observation, cells have their favorite ECM to attach, so perhaps poly-lysine or collagen don't satisfy your cells (this happened to my cells). Maybe you can try coating the muti-well chamber with other ECMs.
Another way you probably won't like it is to synchronize the cells and stain them after the fixation at different time points.
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