TMRM Perchlorate  (Synonyms: T668)

Rhodamine dyes are membrane-permeable cationic fluorescent probes that specifically recognize mitochondrial membrane potentials, thereby attaching to mitochondria and producing bright fluorescence, and at certain concentrations, rhodamine dyes have low toxicity to cells, so they are commonly used to detect mitochondria in animal cells, plant cells, and microorganisms^[1].

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 TMRM working solution
1.1 Preparation of storage solution
Prepare a 5 mM solution by diluting TMRM with anhydrous DMSO.
1.2 Preparation of Working Solution
Dilute the stored solution with preheated serum-free cell culture medium or PBS to prepare a TMRM working solution of 1-20 μM.
Note: Please adjust the concentration of the TMRM working solution according to the actual situation, and prepare it as needed.
2. Cell Staining (Suspension Cells)
2.1 At 4℃, perform a 1000 g centrifugation for 3-5 minutes to collect the cells. Add PBS for two washes, each for 5 minutes. The cell density is 1×10⁶ per mL.
2.2 Add 1 mL of TMRM working solution and incubate at room temperature for 5 to 30 minutes. 2.3 At 4°C, centrifuge at 400 g for 3 to 4 minutes, and 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. Cell Staining (Monolayer Cells)
3.1 Place the monolayer cells on a sterile cover slip.
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 5 to 30 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.

500.93

C₂₅H₂₅ClN₂O₇

576

550

115532-50-8

Solid

Green to dark green

O=C(C1=CC=CC=C1C2=C3C=CC(N(C)C)=CC3=[O+]C4=C2C=CC(N(C)C)=C4)OC.O=Cl(=O)([O-])=O

Room temperature in continental US; may vary elsewhere.

4°C, sealed storage, away from moisture and light

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

* 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, 2 years; -20°C, 1 year (sealed storage, away from moisture and light). When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

• [1]. Crowley LC, et al. Measuring Mitochondrial Transmembrane Potential by TMRE Staining. Cold Spring Harb Protoc. 2016 Dec 1;2016(12):pdb.prot087361.  [Content Brief]

  [2]. Chowdhury SR, et al. Simultaneous evaluation of substrate-dependent oxygen consumption rates and mitochondrial membrane potential by TMRM and safranin in cortical mitochondria. Biosci Rep. 2015 Dec 8;36(1):e00286.  [Content Brief]

  [3]. Monteith A, et al. Imaging of mitochondrial and non-mitochondrial responses in cultured rat hippocampal neurons exposed to micromolar concentrations of TMRM. PLoS One. 2013;8(3):e58059.  [Content Brief]