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Mitotracker green fm probe

Manufactured by Thermo Fisher Scientific
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The MitoTracker Green FM probe is a fluorescent dye that selectively stains mitochondria in live cells. It is a lipophilic, cationic dye that accumulates in active mitochondria as a result of their negative membrane potential.

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Lab products found in correlation

Lsm 710 microscope, by Zeiss (2 mentions) Mitosox reagent, by Thermo Fisher Scientific (2 mentions) Upright fluorescence confocal microscope, by Olympus (1 mentions) Spectramax i3, by Molecular Devices (1 mentions) Hoechst 33342, by Thermo Fisher Scientific (1 mentions) Mitotracker green fm, by Thermo Fisher Scientific (1 mentions) Lysotracker deep red, by Thermo Fisher Scientific (1 mentions) Hoechst 33342, by Merck Group (1 mentions) Bodipy 558 568 c12, by Thermo Fisher Scientific (1 mentions) Synergy h1 reader, by Agilent Technologies (1 mentions) Accuri c6, by BD (1 mentions) Lsm 800, by Zeiss (1 mentions) Facscanto 2, by BD (1 mentions) Facsdiva, by BD (1 mentions) Tmre dye, by Abcam (1 mentions) Lsrfortessa, by BD (1 mentions) Sytox blue dead stain, by Thermo Fisher Scientific (1 mentions) Mitoview 633, by Biotium (1 mentions) Dulbecco phosphate buffered saline (dpbs), by Thermo Fisher Scientific (1 mentions) Trypsin, by Merck Group (1 mentions)

Close spelling variants of this product

MitoTracker Green (911 citations) MitoTracker Green FM (851 citations) MitoTracker (469 citations) MitoTracker probe (48 citations) Mitotracker Green probe (10 citations) MitoTracker green FM fluorescent probe (2 citations)

16 protocols using mitotracker green fm probe

1

Analyzing Mitochondrial Morphology in HCC Cells

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The mitochondrial morphology in the HCC cells was observed by staining with a MitoTracker Green FM probe (Invitrogen) in the dark for 30 min at 37℃. Images were obtained under a Zeiss 880 confocal microscope to assess the number and morphology of mitochondria. Finally, the percentage of cells with small and round mitochondria was calculated.
Zheng J., Yan X., Lu T., Song W., Li Y., Liang J., Zhang J., Cai J., Sui X., Xiao J., Chen H., Chen G., Zhang Q., Liu Y., Yang Y., Zheng K, & Pan Z. (2023). CircFOXK2 promotes hepatocellular carcinoma progression and leads to a poor clinical prognosis via regulating the Warburg effect. Journal of Experimental & Clinical Cancer Research : CR, 42, 63.
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2

Mitochondrial Distribution in Treated Cells

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To analyze mitochondrial distribution, treated-PCCN cells were incubated with pre-warmed (37°C) staining solution containing the Mitotracker Green FM probe (M7514) (Invitrogen) (final concentration 100 nM) for 45 min. After staining is complete, the staining solution was replaced with prewarmed fresh media and observed using an upright fluorescence confocal microscope (Olympus, Tokyo, Japan). The number of cells with non-tubular/fragmented mitochondria was determined, with at least 500 transfected cells examined. All quantifications were based on at least three independent experiments.
Song Z., Shah S.Z., Yang W., Dong H., Yang L., Zhou X, & Zhao D. (2017). Downregulation of the Repressor Element 1-Silencing Transcription Factor (REST) Is Associated with Akt-mTOR and Wnt-β-Catenin Signaling in Prion Diseases Models. Frontiers in Molecular Neuroscience, 10, 128.
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3

Visualizing Mitochondrial Morphology

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Mitochondrial morphology was visualized by MitoTracker Green FM staining using a confocal microscopy. Briefly, HCC cells were collected and seeded to a confocal dish, and cultured overnight. Subsequently, cells were fixed in 4% paraformaldehyde (PFA; Sigma) for 30 min and incubated with a MitoTracker green FM probe (Molecular Probes, M7514) for 1.5 h. The immunofluorescence images were taken with an Olympus FV 1000 laser-scanning confocal microscope (Olympus Corporation, Tokyo, Japan). The length of mitochondria was measured with the ImageJ software (NIH, Bethesda, MD).
Zhang Y., Li H., Chang H., Du L., Hai J., Geng X, & Yan X. (2018). MTP18 overexpression contributes to tumor growth and metastasis and associates with poor survival in hepatocellular carcinoma. Cell Death & Disease, 9(10), 956.
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4

Mitochondrial Staining in MM.1S Cells

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Mitochondria were stained by incubating live MM.1S and MM.1S BzR cells with the MitoTracker Green FM probe (Molecular Probes, 200 nM) according to the manufacturer's instructions. Cells were washed and seeded in triplicate at varying cell densities in 200 μL of growth medium. Fluorescence was quantified using a Spectramax i3 multimode microplate reader (Molecular Devices; Ex: 490 nm, Em: 516 nm). Alternatively, cells were incubated with a dose range of the MitoTracker Green FM probe and a fixed number of cells (0.2 × 106) were plated. Fluorescence intensity was measured as described above.
Thompson R.M., Dytfeld D., Reyes L., Robinson R.M., Smith B., Manevich Y., Jakubowiak A., Komarnicki M., Przybylowicz-Chalecka A., Szczepaniak T., Mitra A.K., Van Ness B.G., Luczak M, & Dolloff N.G. (2017). Glutaminase inhibitor CB-839 synergizes with carfilzomib in resistant multiple myeloma cells. Oncotarget, 8(22), 35863-35876.
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5

Mitochondrial Morphology Visualization

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Mitochondrial morphology was measured with 20 nM MitoTracker Green FM probe (Molecular Probes, Invitrogen, UK), according to the manufacturer's instructions. Briefly, cells were stained with MitoTracker Green for 30 min and Hoechst (1 μg/ml) for 15 min at 37°C.
Zheng Z., Tang D., Zhao L., Li W., Han J., Hu B., Nie G, & He Y. (2020). Liproxstatin-1 Protects Hair Cell-Like HEI-OC1 Cells and Cochlear Hair Cells against Neomycin Ototoxicity. Oxidative Medicine and Cellular Longevity, 2020, 1782659.
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6

Mitochondrial Imaging and Analysis

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To label the mitochondria, cells were incubated with 100mM MitoTracker Green FM probe (Thermo Fisher Scientific; M7514) at 37°C for 30mins. Cells were imaged at 63× oil using Zeiss LSM710 microscope.
For mitochondrial fragmentation measurements, cells were incubated in control or no (0%) glucose medium for 16 h. Cells were then treated with mitochondrial dye along with 100mM LysoTracker DeepRed (ThermoFisher Scientific; L12492) and Hoechst33342 (Sigma-Aldrich; B2261). Mitochondrial fragmentation was calculated in ImageJ using a modified version of MiNA plug-in to threshold for fragment length.
For membrane potential measurements cell were treated with 100 μM TMRE (ThermoFisher Scientific, T669) along with MitoTracker Green FM and Hoechst33342 for 30mins at 37°C. TMRE:MitoTracker ratio was calculated in ImageJ using the MitoMorphology macro.
For lipid trafficking into the mitochondria, MEFs were incubated with 1μM BODIPY 558/568 C12 (Thermo Fisher Scientific; D3835) overnight and then chased in control medium or HBSS for 16 hours (Rambold et al., 2015 (link)). The cells were then stained with the mitochondrial dye and Hoechst33342 for 30mins at 37°C and imaged. Colocalization was calculated in ImageJ using the colocalization plug-in JACoP for the lipid trafficking experiment.
Ravi A., Palamiuc L., Loughran R.M., Triscott J., Arora G.K., Kumar A., Tieu V., Pauli C., Reist M., Lew R.J., Houlihan S.L., Fellmann C., Metallo C., Rubin M.A, & Emerling B.M. (2021). PI5P4Ks drive metabolic homeostasis through peroxisome-mitochondria interplay. Developmental cell, 56(11), 1661-1676.e10.
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7

Mitochondrial Membrane Potential Assay

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Cells were plated on a 24‐well plate at the density of 1 × 105. After 24 h of treatment, cells were washed with serum‐free medium and loaded with MitoTracker Green FM probe (Thermo Fisher Scientific) at a final concentration of 100 nm in a well for 30 min at 37 °C and 5% CO2 in the dark. Afterward, cells were washed twice with 1 × PBS. Fluorescence was measured using a Synergy H1 Reader (BioTek) at an excitation of 490 nm and an emission of 516 nm.
Liskova V., Kajsik M., Chovancova B., Roller L, & Krizanova O. (2022). Camptothecin, triptolide, and apoptosis inducer kit have differential effects on mitochondria in colorectal carcinoma cells. FEBS Open Bio, 12(5), 913-924.
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8

Mitochondrial ROS Measurement by Flow

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Cells were incubated with 5 μM MitoSOX reagent (Thermo Fisher Scientific, M36008) or 25–100 nM MitoTracker Green FM probe (Thermo Fisher Scientific, M7514) at 37 °C for 30 min. After staining was complete, the cells were gently washed three times with warm PBS. Detach cells from plates and ensure a single cell suspension. Analyze the samples by flow cytometry using BD Accuri C6.
Huangyang P., Li F., Lee P., Nissim I., Weljie A.M., Mancuso A., Li B., Keith B., Yoon S.S, & Simon M.C. (2019). Fructose-1,6-bisphosphatase 2 inhibits sarcoma progression by restraining mitochondrial biogenesis. Cell metabolism, 31(1), 174-188.e7.
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9

Mitochondrial Membrane Potential Assay

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Cells were incubated with 50 nM MitoTracker Green FM probe (Thermo Fisher Scientific, M7514) or 5 μM MitoSOX reagent (Thermo Fisher Scientific, M36008) at 37 °C for 30 min. Cells were then washed and analyzed by flow cytometry (Excitation wavelength 488 nm and 594 nm).
Zhao Q., Zhang C., Zhang X., Wang S., Guo T., Yin Y., Zhang H., Li Z., Si Y., Lu Y., Cheng S, & Ding W. (2023). ZNF281 inhibits mitochondrial biogenesis to facilitate metastasis of hepatocellular carcinoma. Cell Death Discovery, 9, 396.
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10

Visualizing Mitochondria and Lysosomes

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Cells were grown on coverslips inside a cell culture dish. After treatment, the culture medium in the dish was replaced with prewarmed staining solution containing 200 nM MitoTracker™ Green FM probe (Thermo Fisher Scientific, USA). Cells were incubated with the staining solution for 30 min at 37 °C. To visualize lysosome, cells were then incubated in a prewarmed staining solution containing 25 nM LysoTracker™ Red DND-99 probe for the next 10 min. After staining, the cells were washed two times with PBS and incubated with fresh media. Then, the cells were visualized using a Zeiss confocal laser scanning microscope (Carl Zeiss, LSM 800). The results were obtained from at least 30 cells in each group.
Yang X., Xue P., Chen H., Yuan M., Kang Y., Duscher D., Machens H.G, & Chen Z. (2020). Denervation drives skeletal muscle atrophy and induces mitochondrial dysfunction, mitophagy and apoptosis via miR-142a-5p/MFN1 axis. Theranostics, 10(3), 1415-1432.
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