1：Experimental Design and Method Selection:

The study involved developing and testing a theranostic pair targeting CD146 in melanoma. Methods included flow cytometry for CD146 expression assessment, synthesis of radiolabeled antibodies for PET imaging, and conjugation with IR700 for photoimmunotherapy. In vitro and in vivo experiments were conducted to evaluate diagnostic and therapeutic efficacy.

2：Sample Selection and Data Sources:

Melanoma cell lines A375 and SK-MEL-5 were used. Subcutaneous tumor models in athymic nude mice were established for in vivo studies. Data were collected from PET imaging, biodistribution studies, fluorescence imaging, and histopathological analyses.

3：List of Experimental Equipment and Materials:

Equipment included flow cytometer (BD LSR Fortessa), PET/CT scanner (Inveon μPET/μCT, Siemens), IVIS Spectrum imaging system (Perkin Elmer), confocal microscope (Nikon A1R), γ-counter (Perkin Elmer), UV-Vis spectrophotometer (Cary 60, Agilent), and laser system (Diode Laser System, Laserglow). Materials included antibodies (YY146, IgG), radionuclides (89Zr, 18F-FDG), dyes (IR700, CellROX Green, Hoechst 33342), cell culture media, and chemicals for conjugation and staining.

4：Experimental Procedures and Operational Workflow:

Procedures involved cell culture and flow cytometry for CD146 expression, radiolabeling of antibodies with 89Zr, PET imaging at various time points, biodistribution studies, synthesis of IR700 conjugates, in vitro PIT with laser irradiation, in vivo PIT in mouse models, fluorescence imaging, and histopathological staining (H&E and immunofluorescence).

5：Data Analysis Methods:

Data were analyzed using FlowJo for flow cytometry, Inveon Research Workplace for PET image reconstruction, Living Image software for fluorescence imaging, and Prism software for statistical analysis (Student's t-tests and ANOVA).