After tagging separate cell population with mitotracker Green and Deep-red, Co-culture at different time points(24hr onwards) and confluency produces same result i.e. 100% transfer/exchange of mitochondria in MDA-MB231 cell line. All of the cell population are in the quadrant positive for Red and Green.
Does the tracker dye diffuse under culture conditions? Con-focal imaging showed overlapping, i.e. yellow colored mitochondria. Mixing different cell population just before flow-cytometry produces separate population in dot plot.
This observation of 100% exchange does not seem normal
Dr. Uhlen, We are still trying to make stable transfected cell line. I also think that somehow the dyes are diffusing. Thank you for your suggestion.
Qdots (QTracker from Life Tech)? I've never used them, but I know they're for long term labeling and shouldn't transfer.
I'm with Molecular Probes Tech Support. Though MitoTrackers have "tracker" in the name, they are not intended for long-term labeling or tracking of mitochondria. They are only for end-point imaging (for mitochondrial localization, morphology, or membrane potential, or mass, depending on which one and which assay). Long term exposure can actually cause mitochondrial toxicity and cell toxicity.
We have other cell tracking dyes, such as our CellTracker products, CFDA SE, or Qtracker Qdots (as Matt mentioned). BUT none of those target mitochondria. If you wish to track mitochondria, then the only way I can think of, in live cells, is to use a fluorescent protein that targets a mitochondrial protein. Our CellLight Mito products can work for that (for mammalian non-hematopoietic cell lines) for up to 3-5 days (it's a transient transfection method).
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