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MitoTracker™ Red FM - Special Packaging, 20 x 50 μg, 20 x 50 μg - FAQs

MitoTracker™ Red FM - Special Packaging, 20 x 50 μg, 20 x 50 μg - FAQs

View additional product information for MitoTracker™ Dyes for Mitochondria Labeling - FAQs (M22426, M22425, M7514, M7512, M7513, M7510, M7511)

5 product FAQs found

It looks like my Mitotracker dye is staining more than just the mitochondria. Why?

This is typically a result of using too high of a concentration of the Mitotracker dye. Most organic dyes are used in the low micromolar range. The MitoTracker dyes are used at a much lower concentration, around 50–200 nanomolar. Higher concentrations can cause background fluorescence and non-mitochondrial staining.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Can I use MitoTracker Red CMXRos dye in a plate reader to study mitochondrial membrane potential changes?

Yes, it has been done. A literature search will find numerous examples. However, be aware that plate readers are typically less sensitive than microscopes or flow cytometers, which means that the degree of change in fluorescence may not be as sensitive or easily detectable with a plate reader. Be sure to optimize label times and concentrations carefully.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Our facility does not allow flow cytometry of live cells. Can I fix the cells after staining with MitoTracker Red FM dye?

No, this wouldn't work as MitoTracker Red FM dye is not retained upon fixation. See Table 2 in this manual (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/mp07510.pdf).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

After I labeled neurons with MitoTracker Red CMXRos, they are dead the next day. Is this expected?

The Mitotracker dyes should be imaged soon after staining because over time, those dyes can be toxic.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I am testing mitochondrial membrane potential, but my untreated cells are fluorescing, and I'm not seeing a significant difference in my test sample.

Regardless of which dye you use - tetramethylrhodamine, methyl ester (TMRM), JC-1 or MitoTracker - untreated cells will fluoresce. It's just that cells with reduced mitochondrial membrane potential will fluoresce less. It is the degree of change which is important. JC-1 dye not only changes intensity, but has a ratiometric spectral change in excitation and emission. It is very important to have an untreated control as well as a positive control treated with a mitochondrial membrane potential destabilizer, such as CCCP or FCCP. Most mitochondrial stains are only for use with live cells, as the signal will not be retained to the same degree with fixation.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.