1：Experimental Design and Method Selection:

The study involved analyzing dimerization constants of dyes in buffer solutions using UV-Vis and fluorescence spectroscopy, and preparing liposomes via thin-film hydration and extrusion for encapsulation. Theoretical models included dimerization equilibrium equations and Arrhenius plots for diffusion activation energy.

2：Sample Selection and Data Sources:

Dyes (indocyanine green, patent blue V, brilliant blue G) were medical grade and used without purification. Liposomes were prepared from soybean lecithin and cholesterol in PBS buffer (pH 7.4).

3：4). List of Experimental Equipment and Materials:

3. List of Experimental Equipment and Materials: UV-Vis spectrophotometer (Cary 100-Varian), spectrofluorimeter (Varian Eclipse), SEM, DLS particle size analyzer (Zetasizer Nano ZS90, Malvern Instruments), ultrasound homogenizer, extrusion filters (100 nm). Materials included EPK130 lecithin, PBS tablets (Sigma Aldrich), dyes from various suppliers.

4：Experimental Procedures and Operational Workflow:

Dye solutions were prepared in PBS, and spectra were recorded. Liposomes were formed by dissolving lipids in chloroform/methanol, drying, hydrating with PBS, sonicating, extruding, and dialyzing. Size, zeta potential, and encapsulation efficiency were measured.

5：Data Analysis Methods:

Dimerization constants were calculated from integral absorbance data. Diffusion coefficients and activation energies were derived from DLS measurements using Stokes-Einstein and Arrhenius equations. Zeta potential data were used to compute surface charge ratios.