MitoTracker Deep Red FM  (Synonyms: MTDR FM)

MitoTracker Deep Red (MTDR) FM fluorescent dye that selectively accumulates in the mitochondrial matrix. MitoTracker Deep Red FM covalently binds mitochondrial proteins by reacting with free mercaptan of cysteine residues, allowing staining of mitochondrial membrane potential independent of membrane potential. Excitation/emission wavelength 644/665 nm^[1]. MitoTracker Deep Red dyes have an excitation/emission wavelength of 633/650-750 nm^[2].
The Ex/Em of MitoTracker Deep Red (MTDR) FM is 644/665 nm.

Guide (The following is the experimental plan we recommend. This plan serves only as a reference guide. The specific operations should be adjusted according to your actual needs.)
1. Preparation of MitoTracker Deep Red FM working solution
1.1 Preparation of storage solution
Prepare a 1 mM solution of MitoTracker Deep Red FM using DMSO.
1.2 Preparation of working solution
Use pre-heated serum-free cell culture medium or PBS to dilute the storage solution at a ratio of 1:5000 - 1:50000, and prepare a 20 - 200 nM MitoTracker Deep Red FM working solution.
Note: Please adjust the concentration of MitoTracker Deep Red FM working solution according to the actual situation, and prepare it as needed.

2. Cell Staining (Suspension Cells)
2.1 Centrifuge to collect the cells, then add PBS for two washes, each for 5 minutes. The cell density is 1×10⁶ per mL.
2.2 Add 1 mL of MitoTracker Deep Red FM solution and incubate at room temperature for 15 to 45 minutes.
2.3 400 grams, centrifuge for 3-4 minutes, then discard the supernatant.
2.4 Add PBS to wash the cells twice, for 5 minutes each time.
2.5 After resuspending the cells in 1 mL of serum-free medium or PBS, observe them using a fluorescence microscope or a flow cytometer.
3. Cytological staining (adherent cells) 3.1 Cultivate adherent cells on a sterile cover glass.
3.2 Remove the cover glass from the culture medium and aspirate away the excess medium.
3.3 Add 100 μL of the dye working solution and gently shake to ensure complete coverage of the cells. Incubate for 15 to 45 minutes.
3.4 Remove the dye working solution and wash with the culture medium 2-3 times, for 5 minutes each time. Then observe using a fluorescence microscope or flow cytometer.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

543.57

C₃₄H₃₆Cl₂N₂

873315-86-7

Solid

Blue to dark blue

ClCC1=CC=C(CN2C3=C(C(C)(C)/C2=C/C=C/C=C/C4=[N+](C)C5=C(C4(C)C)C=CC=C5)C=CC=C3)C=C1.[Cl-]

Room temperature in continental US; may vary elsewhere.

-20°C, protect from light, stored under nitrogen

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen)

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

• [1]. Buckman JF, et al. MitoTracker labeling in primary neuronal and astrocytic cultures: influence of mitochondrial membrane potential and oxidants. J Neurosci Methods. 2001 Jan 15;104(2):165-76.  [Content Brief]

  [2]. Wang FX, et al. Mitochondria-Accumulating Rhenium(I) Tricarbonyl Complexes Induce Cell Death via Irreversible Oxidative Stress and Glutathione Metabolism Disturbance. ACS Appl Mater Interfaces. 2019 Apr 10;11(14):13123-13133.  [Content Brief]