Objectives: The aim of this study was to formulate fluorescent-labeled targeted immunoliposome to visualize the delivery and distribution of drugs in real-time.

Methods: In this study, fluorescent-labeled liposomes were decorated with anti-HER2 VHH or Herceptin to improve the monitoring of intracellular drug delivery and tumor cell tracking with minimal side effects. The conjugation efficiency of antibodies was analyzed by SDS-PAGE silver staining. In addition, the physicochemical characterization of liposomes was performed using DLS and TEM. Finally, confocal microscopy visualized nanoparticles in the target cells.

Results: Quantitative and qualitative methods characterized the intracellular uptake of 110±10 nm particles with near 70% conjugation efficiency. In addition, live-cell trafficking during hours of incubation was monitored by wide-field microscopy imaging. The results show that the fluorescent- labeled nanoparticles can specifically bind to HER2-positive breast cancer with minimal off-target delivery.

Conclusion: These nanoparticles can have several applications in personalized medicine, especially drug delivery and real-time visualization of cancer therapy. Moreover, this method also can be applied in the targeted delivery of contrast agents in imaging and thermotherapy.

目的：本研究的目的是制定荧光标记的靶向免疫脂质体，以实时可视化药物的递送和分布。

方法：在这项研究中，荧光标记的脂质体用抗 HER2 VHH 或赫赛汀修饰，以改善对细胞内药物递送和肿瘤细胞追踪的监测，并且副作用最小。通过SDS-PAGE银染分析抗体的缀合效率。此外，使用 DLS 和 TEM 进行脂质体的物理化学表征。最后，共聚焦显微镜观察了靶细胞中的纳米颗粒。

结果：定量和定性方法表征了 110±10 nm 颗粒的细胞内吸收，结合效率接近 70%。此外，通过广角显微镜成像监测孵化数小时内的活细胞运输。结果表明，荧光标记的纳米颗粒可以特异性结合 HER2 阳性乳腺癌，并且具有最小的脱靶递送。

结论：这些纳米颗粒可以在个性化医疗中具有多种应用，尤其是药物输送和癌症治疗的实时可视化。此外，该方法还可应用于成像和热疗中造影剂的靶向输送。