7. Monitoring changes in copper ion concentration

Cells suspended in MEM were treated with 20 µM Phen Green FL, a fluorescent copper indicator, for 15 minutes. After dye loading, cells were washed three times with PBS. After treatment with 20 µM fluoxetine alone, or in combination with bathocuproine disulfonic acid and deferoxamine, the fluorescence of Phen Green FL was measured at an excitation wavelength of 492 nm and an emission wavelength of 517 nm using a fluorescent microscope.