1：Experimental Design and Method Selection:

The study involved synthesizing DCP-Cy from IR-820 via Suzuki coupling, characterizing its solubility and spectral properties, encapsulating it in liposomes, and evaluating its performance in vitro and in vivo using fluorescence and photoacoustic imaging. Theoretical models include J-aggregate formation kinetics and liposome formulation principles.

2：Sample Selection and Data Sources:

DCP-Cy was synthesized and tested in aqueous solutions with varying pH, salt concentrations, and in liposomes. In vivo studies used BALB/c and CD-1 mice for pharmacokinetics, biodistribution, and imaging. Data were collected from spectral analyses, dynamic light scattering, cryo-electron microscopy, and imaging systems.

3：List of Experimental Equipment and Materials:

Equipment includes Perkin Elmer Lambda 35 UV/VIS Spectrometer, NanoBrook 90 plus PALS for DLS, Tecnai F20 electron microscope for cryo-EM, Verasonics Vantage system for photoacoustic imaging, custom fluorescence imaging setup, and homogenizers. Materials include IR-820, 3,5-Dicarboxyphenylboronic acid, lipids (DOPC, DSPC, cholesterol, DSPE-PEG), salts, buffers, and solvents.

4：Experimental Procedures and Operational Workflow:

Synthesis of DCP-Cy, solubility tests, spectral characterization, liposome preparation via ethanol injection method, size exclusion chromatography, in vitro tests with detergents, in vivo administration in mice, blood collection, organ homogenization, and imaging with photoacoustic and fluorescence systems.

5：Data Analysis Methods:

Data were analyzed using absorbance ratios (934 nm/789 nm), standard curves for quantification, ImageJ for fluorescence intensity, and back-projection algorithms for photoacoustic image reconstruction. Statistical analyses included mean calculations for pharmacokinetics and biodistribution.