1：Experimental Design and Method Selection:

The study designed a phospholipid nanoprobe (PL(I/D)NP) loaded with Lipiodol and DiR-BOA for dual-modality imaging. The experimental design involved optimizing nanoparticle formulations for size and stability, characterizing in vitro properties, and evaluating in vivo lymphatic tracing and metastasis diagnosis using micro-CT and fluorescence imaging. Theoretical models included nanoparticle size optimization for lymphatic drainage and SR-B1 receptor targeting.

2：Sample Selection and Data Sources:

Mouse models (C57BL/6 and BALB/c) were used for in vivo studies. Cell lines included 4T1 murine mammary adenocarcinoma cells, ldlA7 cells, and ldlA(mSR-B1) cells. Samples were selected based on standard animal and cell culture protocols.

3：List of Experimental Equipment and Materials:

Equipment included Zetasizer Nano-ZS90 for DLS, transmission electron microscope (TECNAI G2), UV2550 UV-vis spectrophotometer, LS55 luminescence spectrometer, custom-made micro-CT system, confocal microscope (Zeiss LSM 710), flow cytometer (Guava easyCyte 8HT), and fluorescence imaging systems. Materials included phospholipids (DMPC, DMPE, MHPC, DSPE-PEG2000), Lipiodol, DiR-BOA, Omnipaque, ICG, and other chemicals.

4：Experimental Procedures and Operational Workflow:

Nanoparticles were prepared via thin-film hydration method, characterized for size, zeta potential, and stability. In vitro imaging and cell uptake studies were performed. In vivo, nanoparticles were injected subcutaneously into mice, and CT/fluorescence imaging was conducted at various time points. Lymph node metastasis models were established with 4T1 cells, and histopathological analysis was done post-imaging.

5：Data Analysis Methods:

Data were analyzed using MATLAB for CT attenuation, GraphPad Prism for statistical analysis (Student's t-test), and flow cytometry/confocal imaging for quantitative uptake. LN volumes and LV lengths were measured from CT images.