Bertorelle Franck, Wilhelm Claire, Roger Jacky, Gazeau Florence, Ménager Christine, Cabuil Valérie
Laboratoire des Liquides Ioniques et Interfaces Chargées, Equipe Colloïdes Inorganiques, UMR 7612 CNRS/Université Pierre et Marie Curie (Paris 6), 4 place Jussieu, case 63, 75252 Paris, Cedex 05, France.
Langmuir. 2006 Jun 6;22(12):5385-91. doi: 10.1021/la052710u.
This report describes the preparation and characterization of new magnetic fluorescent nanoparticles and our success in using them to label living cells. The bifunctional nanoparticles possess a magnetic oxide core composed of a dimercaptosuccinic acid (DMSA) ligand at the surface and a covalently attached fluorescent dye. The nanoparticles exhibited a high affinity for cells, which was demonstrated by fluorescence microscopy and magnetophoresis. Fluorescence microscopy was used to monitor the localization patterns of magnetic nanoparticles associated with cells. We observed two types of magnetic labeling: adsorption of the nanoparticles on the cell membrane (membranous fluorescence) and internalization of the nanoparticles inside the cell (intracellular vesicular fluorescence). After internalization, nanoparticles were confined inside endosomes, which are submicrometric vesicles of the endocytotic pathway. We demonstrated that endosome movement could be piloted inside the cell by external magnetic fields such that small fluorescent chains of magnetic endosomes were formed in the cell cytoplasm in the direction of the applied magnetic field. Finally, by measuring the critical cellular magnetic load (quantitated by magnetophoresis), we have demonstrated the potential of this new magneto-fluorescent nanoagent for medical use.
本报告描述了新型磁性荧光纳米颗粒的制备与表征,以及我们成功使用它们标记活细胞的过程。这种双功能纳米颗粒具有一个磁性氧化物核心,表面由二巯基丁二酸(DMSA)配体组成,并共价连接有荧光染料。纳米颗粒对细胞表现出高亲和力,这通过荧光显微镜和磁泳得以证明。荧光显微镜用于监测与细胞相关的磁性纳米颗粒的定位模式。我们观察到两种类型的磁性标记:纳米颗粒吸附在细胞膜上(膜荧光)和纳米颗粒内化到细胞内部(细胞内囊泡荧光)。内化后,纳米颗粒被限制在内体中,内体是内吞途径的亚微米级囊泡。我们证明,外部磁场可以引导内体在细胞内移动,从而在细胞质中沿外加磁场方向形成磁性内体的小荧光链。最后,通过测量临界细胞磁性负载(通过磁泳定量),我们证明了这种新型磁荧光纳米试剂在医学应用中的潜力。