JC-10 is a potent inhibitor of metalloproteinases and is often used as a probe.

Instructions
1. Solution preparation
1. Stock solution preparation: Dissolve JC-10 dye in anhydrous DMSO to prepare a 100× stock solution.
2. Working solution preparation: Dilute the stock solution to the working concentration using preheated cell culture medium. For example, add 10 µL of JC-10 stock solution to 1 mL of preheated culture medium and mix until the solution changes from pink to nearly colorless.
2. Operation steps
Cell staining
1. Add the dye working solution to the cells, ensuring that the cells are completely immersed.
2. Incubate at 37°C in the dark for 15-30 minutes.
3. Washing: After incubation, remove the dye working solution.
4. Wash the cells 2-3 times with preheated PBS or culture medium to remove unbound dye.
5. Detection: Detect using a fluorescence microscope or flow cytometer.
JC-10 forms aggregates in the mitochondria of healthy cells, showing orange-red fluorescence (excitation/emission wavelength: 540/590 nm); when the membrane potential decreases, JC-10 exists in monomeric form, showing green fluorescence (excitation/emission wavelength: 490/525 nm).
Notes:
1) Wear gloves during operation to avoid contact between skin or mucous membranes and reagents.
2) Avoid light during incubation and storage to prevent fluorescence quenching.
3) After staining, fluorescence detection analysis should be performed as soon as possible.