Jc 1 cationic dye

After exposure to MNPs (0, 10, 30, and 60 μg/ml) for 24 h, MMP in SH-SY5Y cells was measured according to Rana et al. [21 (link)]. MMP were measured using the JC-1 cationic dye (Molecular Probes). SH-SY5Y cells were seeded in 96-well (black with the clear flat bottom) plate and cultured for one day. The cells were exposed to different concentration of MNPs for 24 h and incubated in Hank's balanced salts solution (HBSS) containing 2 μg/ml JC-1 for 15 min. Green and red fluorescence was measured at 450 nm and 570 nm using a microplate reader, respectively. Depolarization of mitochondrial is assessed by the decline in ratio of red/green fluorescence intensity. For qualitative analysis of MMP, cell image was captured using fluorescence by upright microscope fluorescence (Nikon Eclipse 80i).

Mitochondrial membrane potential (Δψm) was monitored utilizing the JC-1 cationic dye (Molecular Probes) as recommended by the manufacturer. HT-29 cells were seeded into 6-well plates (5 × 10⁵ cells/well) and cultured overnight prior to exposure to different concentrations of DCMPI (30 μg/mL, 60 μg/mL) for 24 h and then resuspended in a buffer solution containing 10 μg/mL JC-1 cationic dye. After 20 min of incubation in the dark at 37°C, the samples were rinsed with PBS to remove unreacted dye. The fluorescence intensity was read with the Guava Easy cytometer within 1 h.