1：Experimental Design and Method Selection:

The study involved synthesizing DAF, characterizing its photophysical properties in various solvents, and applying it in cellular imaging to stain lipid droplets and cytoplasm. Ratiometric imaging was used for polarity mapping.

2：Sample Selection and Data Sources:

KB cells (HeLa cell derivatives) were used for cellular imaging. Solvents of varying polarity were used for spectroscopic studies.

3：List of Experimental Equipment and Materials:

Instruments included a Bruker Avance III 400 MHz spectrometer for NMR, Agilent Q-TOF 6520 mass spectrometer for mass spectra, Cary 400 Scan spectrophotometer for absorption, FluoroMax-4 spectrofluorometer for emission, time-correlated single-photon counting setup for lifetime measurements, confocal microscope for imaging, and various chemicals from suppliers like Alfa Aesar, Sigma Aldrich, TCI Europe.

4：Experimental Procedures and Operational Workflow:

Synthesis of DAF via Suzuki coupling, spectroscopy in solvents, cytotoxicity assay using MTT, confocal imaging of live and fixed cells with DAF and other dyes, colocalization studies, and ratiometric imaging.

5：Data Analysis Methods:

Data analyzed using Matlab software for lifetime, Icy software for image processing, SODA plugin for colocalization, and ImageJ for ratiometric images.