Iron Ion Concentration Measurement in AsPC-1 Cells Using Phen Green SK Fluorescence Indicator

Phen Green SK, a fluorescent indicator, was utilized to measure the concentration of iron ions in AsPC-1 cells. The cells were washed thrice with 1 mL of chilled PBS for 1 minute each time, fixed with 4% PFA for 20 minutes, and washed again with cold PBS. After standing in the dark at room temperature for 1 minute, the cells were treated with 10μg/mL of Phen Green SK probe and incubated for 30 minutes at room temperature in the absence of light. Subsequently, the cells were washed thrice with PBS for 1 minute each time and treated with 0.2μg/mL of DAPI solution for 20 minutes, followed by washing three times with chilled PBS. The cells were then mounted upside down on slides with a solution containing 10μL of anti-fluorescence quenching sealing solution and left to stand in the dark for 15 minutes at room temperature. Confocal laser scanning microscopy was used to capture images of the cells, with DAPI being excited at a wavelength of 358 nm and emitting at 461 nm, and Phen Green SK being excited at 488 nm and emitting at 590 nm.