TMRE (Tetramethylrhodamine ethyl ester perchlorate) is a mitochondria specific dye (λex: 550 nm, λem: 575 nm).

Multidirectional dynamic movement of TMRE is observed in epithelial cells and bidirectional dynamic movement is seen in the superficial cortical fiber cells of live bovine lenses. In the epithelium, the movement of TMRE fluorescence is up to 5 μm/min whereas in the superficial cortex the observed movement is up to 18.5 μm/min. The movement of TMRE fluorescence is abolished with the treatment of the uncoupler, carbonyl cyanide m-chlorophenylhydrazone [2].

Instructions for use
I. Solution preparation
1. Preparation of stock solution: Dissolve 1 mg TMRE in DMSO to obtain 5 mM stock solution; (It is recommended to store at -20 ℃ or -80 ℃ in the dark after aliquoting)
2. Preparation of working solution: Dilute the stock solution with serum-free cell culture medium or PBS to obtain a working solution with a final concentration of 1-20 μM. (Select the appropriate working solution concentration according to experimental requirements and prepare it for immediate use)
II. Operation steps
1. Cell staining
Suspended cells (6-well plate)
1) Centrifuge at 1000 g for 3-5 minutes at 4℃, then discard the supernatant. Wash twice with PBS for 5 minutes each. The cell density is 1×106/mL.
2) Add 1 mL of working solution and incubate at room temperature for 5-30 minutes.
3) Centrifuge at 400 g for 3-4 minutes at 4℃, and discard the supernatant.
4) Wash twice with PBS, 5 minutes each time.
5) Resuspend cells with serum-free cell culture medium or PBS. Observe with fluorescence microscope or flow cytometer.
2. Adherent cells
1) Culture adherent cells on sterile coverslips.
2) Remove coverslips from culture medium and aspirate excess culture medium.
3) Add 100 μL working solution, shake gently to completely cover cells, and incubate at room temperature for 30-60 minutes.
4) Wash twice with culture medium, 5 minutes each time. Observe with fluorescence microscope or flow cytometer.