MitoLite™ Green FM
Overview | ![]() ![]() |
See also: Mitochondria, Cell Metabolism, Cell Structures and Organelles, Membrane Potential and Channels
CAS 201860-17-5 | Molecular weight 671.88 | Excitation (nm) 508 | Emission (nm) 528 |
MitoLite™ Green FM is the same molecule to the MitoTracker Green FM (M7514, ThermoFisher). It is green-fluorescent mitochondrial stain. Unlike other MitoLite probes, MitoLite™ Green FM appears to localize to mitochondria, much less depending on mitochondrial membrane potential. The dye stains live cells, but it is not well-retained after aldehyde fixation.
Platform
Flow cytometer
Excitation | 488 nm laser |
Emission | 530/30 nm filter |
Instrument specification(s) | FITC channel |
Fluorescence microscope
Excitation | FITC filter set |
Emission | FITC filter set |
Recommended plate | Black wall/clear bottom |
Example protocol
AT A GLANCE
Protocol Summary
- Prepare 1 mM MitoLite™ Green FM stock solution
- Prepare 20-200 nM MitoLite™ Green FM staining solution
- Remove the growth media from the cells
- Add MitoLite™ Green FM staining solution to cells
- Incubate at 37°C for 30 minutes
- Wash cells and replace with 1x Hanks and 20mM Hepes Buffer (HH buffer)
- Observe cells using a fluorescence microscope with FITC filter set
Important Bring the dye at room temperature before starting the experiment.
PREPARATION OF STOCK SOLUTION
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.
MitoLite™ Green FM stock solution:
Dissolve one vial MitoLite™ Green FM (50 ug) in 74 uL high-quality, anhydrous dimethylsulfoxide (DMSO) to make 1 mM stock solution. Note: Keep the stock solution frozen at ≤–15°C and protected from light.
PREPARATION OF WORKING SOLUTION
MitoLite™ Green FM staining solution:
Dilute 1 mM MitoLite™ Green FM stock solution to the final working concentration in HH buffer. The working concentration can be in the range of 20–200 nM.
SAMPLE EXPERIMENTAL PROTOCOL
Staining adherent cells:
- Grow cells to reach the desired confluency.
- Remove the growth media from the cells.
- Add MitoLite™ Green FM staining solution to each well.
- Incubate at 37°C for 30 minutes.
- Wash cells and replace with 1x Hanks and 20mM Hepes Buffer (HH buffer).
- Observe cells using a fluorescence microscope with FITC filter set. Note: The staining protocols is good for Hela cell line and it may need to be optimized with the particular cell types.
Staining suspension cells:
- Centrifuge cells to a pellet and aspirate the supernatant.
- Resuspend the cells gently in MitoLite™ Green FM staining solution.
- Incubate at 37°C for 30 minutes.
- Centrifuge the cells, remove supernatant and resuspend cells in fresh HH buffer.
- Cells may be analyzed by flow cytometry (530/30 nm filter- FITC channel) or fluorescence microscopy (FITC filter set).
Calculators
Common stock solution preparation
Table 1. Volume of DMSO needed to reconstitute specific mass of MitoLite™ Green FM to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 148.836 µL | 744.181 µL | 1.488 mL | 7.442 mL | 14.884 mL |
5 mM | 29.767 µL | 148.836 µL | 297.672 µL | 1.488 mL | 2.977 mL |
10 mM | 14.884 µL | 74.418 µL | 148.836 µL | 744.181 µL | 1.488 mL |
Molarity calculator
Enter any two values (mass, volume, concentration) to calculate the third.
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
/ | = | x | = |
Citations
View all 1 citations: Citation Explorer
Sonodynamic Therapy Promotes Efferocytosis via CD47 Down-Regulation in Advanced Atherosclerotic Plaque
Authors: Cao, Yang and Yao, Jianting and Gao, Weiwei and Cao, Zhengyu and Diabakte, Kamal and Wang, Linxin and Sun, Xin and Tian, Ye
Journal: International Heart Journal (2022): 21--233
Authors: Cao, Yang and Yao, Jianting and Gao, Weiwei and Cao, Zhengyu and Diabakte, Kamal and Wang, Linxin and Sun, Xin and Tian, Ye
Journal: International Heart Journal (2022): 21--233
References
View all 5 references: Citation Explorer
the influence of platelets and a comparison of analytical approaches
Authors: Hopkinson, K.; Williams, E. A.; Fairburn, B.; Forster, S.; Flower, D. J.; Saxton, J. M.; Pockley, A. G., A MitoTracker Green-based flow cytometric assay for natural killer cell activity: variability
Journal: Exp Hematol (2007): 350-7
Authors: Hopkinson, K.; Williams, E. A.; Fairburn, B.; Forster, S.; Flower, D. J.; Saxton, J. M.; Pockley, A. G., A MitoTracker Green-based flow cytometric assay for natural killer cell activity: variability
Journal: Exp Hematol (2007): 350-7
Efficacy of MitoTracker Green and CMXrosamine to measure changes in mitochondrial membrane potentials in living cells and tissues
Authors: Pendergrass, W.; Wolf, N.; Poot, M.
Journal: Cytometry A (2004): 162-9
Authors: Pendergrass, W.; Wolf, N.; Poot, M.
Journal: Cytometry A (2004): 162-9
MitoTracker Green labeling of mitochondrial proteins and their subsequent analysis by capillary electrophoresis with laser-induced fluorescence detection
Authors: Presley, A. D.; Fuller, K. M.; Arriaga, E. A.
Journal: J Chromatogr B Analyt Technol Biomed Life Sci (2003): 141-50
Authors: Presley, A. D.; Fuller, K. M.; Arriaga, E. A.
Journal: J Chromatogr B Analyt Technol Biomed Life Sci (2003): 141-50
Mitotracker green is a P-glycoprotein substrate
Authors: Marques-Santos, L. F.; Oliveira, J. G.; Maia, R. C.; Rumjanek, V. M.
Journal: Biosci Rep (2003): 199-212
Authors: Marques-Santos, L. F.; Oliveira, J. G.; Maia, R. C.; Rumjanek, V. M.
Journal: Biosci Rep (2003): 199-212
and MitoTracker Green is affected by mitochondrial membrane potential altering drugs
Authors: Keij, J. F.; Bell-Prince, C.; Steinkamp, J. A., Staining of mitochondrial membranes with 10-nonyl acridine orange, MitoFluor Green
Journal: Cytometry (2000): 203-10
Authors: Keij, J. F.; Bell-Prince, C.; Steinkamp, J. A., Staining of mitochondrial membranes with 10-nonyl acridine orange, MitoFluor Green
Journal: Cytometry (2000): 203-10
Application notes
A Novel Fluorescent Probe for Imaging and Detecting Hydroxyl Radical in Living Cells
Activation of the Mitochondrial Apoptotic Pathway Produces Reactive Oxygen Species and Oxidative Damage in Hepatocytes That Contribute to Liver Tumorigenesis
Calmodulin antagonists induce cell cycle arrest and apoptosis in vitro and inhibit tumor growth in vivo in human multiple myeloma
Cold exposure lowers energy expenditure at the cellular level
Developing high-content assays for hepatoxicity with iPSC-derived cells