Rhodamine 123 (solution) 

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].
Solution Concentration: 5 mM

1.Preparation of Rhodamine 123 working solution
1.1Preparation of the stock solution
Dissolve 1 mg Rhodamine 123 in 525 μL DMSO to obtain 5 mM of stock solution.
1.2Preparation of Rhodamine 123 working solution
Dilute the stock solution in serum-free cell culture medium. The corresponding stock solution can be diluted according to the actual situation. Note that if the solvent is DMSO, the cytotoxicity of DMSO must be considered, and a solvent control should be prepared; if the solvent is pure water, the working solution needs to be filtered and sterilized before adding cells.
Note: Please adjust the concentration of Rhodamine 123 working solution according to the actual situation.
2.Cell staining
2.1 Suspension cells (6-well plate)
a.Centrifuge at 1000 g at 4℃ for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.The cell density is 1×10⁶/mL.
b.Add 1 mL of working solution, and then incubate at room temperature for 5-30 minutes.
c.Centrifuge at 400 g at 4℃ for 3-4 minutes and then discard the supernatant.
d.Wash twice with PBS, 5 minutes each time.
e.Resuspend cells with serum-free cell culture medium or PBS. Observation by fluorescence microscopy or flow cytometry.
2.2 Adherent cells
a.Culture adherent cells on sterile coverslips.
b.Remove the coverslip from the medium and aspirate excess medium.
c.Add 100 μL of working solution, gently shake it to completely cover the cells,and then incubate at room temperature for 30-60 minutes.
d.Wash twice with medium, 5 minutes each time. Observation by fluorescence microscopy or flow cytometry.
Note: If detection by flow cytometry, cells need to be resuspended before staining.

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

380.83

C₂₁H₁₇ClN₂O₃

62669-70-9

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

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• [1]. M. Huang, et al. Mitochondrial Inner Membrane Electrophysiology Assessed by Rhodamine-123 Transport and Fluorescence. Ann Biomed Eng. 2007 Jul; 35(7): 1276–1285.  [Content Brief]

  [2]. Emaus, R. K., Grunwald, R., & Lemasters, J. J. (1986). Rhodamine 123 as a probe of transmembrane potential in isolated rat-liver mitochondria: spectral and metabolic properties. Biochimica et Biophysica Acta (BBA) - Bioenergetics, 850(3), 436–448.