This chemical is widely used as a fluorescent pH indicator in biological research. Its ability to exhibit distinct fluorescence changes with varying pH levels makes it particularly valuable for studying intracellular pH dynamics. Researchers often employ it to monitor pH fluctuations in live cells, as it can easily penetrate cell membranes due to its acetoxymethyl ester form, which is then hydrolyzed intracellularly to release the active dye. It is commonly applied in fluorescence microscopy and flow cytometry to investigate cellular processes such as acidification, metabolic activity, and ion transport. Its dual-emission properties allow for ratiometric measurements, enhancing accuracy and minimizing artifacts caused by variations in dye concentration or photobleaching. This makes it a reliable tool for studying pH-related physiological and pathological conditions in various cell types.