Confocal immunofluorescent analysis of HeLa cells using MitoTracker® Green FM. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Each vial of MitoTracker® Green FM contains 50 µg lyophilized solid. To make a 1 mM stock solution, reconstitute the solid in 74.4 µl high quality DMSO. While the optimal staining concentration and incubation times can vary by application, typical results are obtained by diluting the stock solution directly into normal growth media at a concentration between 100-400 nM and incubating between 15-30 min at 37ºC. After incubation, the cells must be imaged live; DO NOT FIX!
To reduce background fluorescence of the media, phenol red free media can be substituted for normal media prior to imaging. BackDrop® Green Background Suppressor #12388 can also be added to reduce background fluorescence.
Store lyophilized solid at -20°C dessicated and protected from light. In lyophilized form, this reagent is stable for 12 months. Once reconstituted in DMSO, the solution should be stored at -20°C, protected from light, and must be used within 2 weeks. Avoid freeze-thaw cycles.
MitoTracker® Green FM is recommended for live cell imaging only; fixation with aldehydes or alcohols will inhibit staining. Excitation: 490 nm, Emission: 516 nm, Molecular Weight: 671.88 g/mol
MitoTracker® dyes are cell permeable probes that contain a mildly thiol-reactive chloromethyl moiety for mitochondrial labeling. To label mitochondria, cells are simply incubated with MitoTracker® probes, which passively diffuse across the plasma membrane and accumulate in active mitochondria.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
BackDrop® is a registered trademark of Molecular Probes, Inc.
DRAQ5 is a registered trademark of Biostatus Limited.
MitoTracker is a registered trademark of Molecular Probes, Inc.