Labeling of immune cells for in vivo imaging using magnetofluorescent nanoparticles
Labeling of immune cells for in vivo imaging using magnetofluorescent nanoparticles
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DOI：
10.1038/nprot.2006.11
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发表时间：
2006-01-01
期刊：
影响因子：
14.8
通讯作者：
Weissleder, Ralph
中科院分区：
文献类型：
--
作者：
Pittet, Mikael J.;Swirski, Filip K.;Weissleder, Ralph
观察免疫细胞和干细胞在其天然微环境中需要开发成像剂以允许其在体内跟踪。我们在这里描述的磁荧光纳米粒子的合成细胞标记在体外和多模态成像的管理细胞在体内。MION-47是一种原型单晶氧化铁纳米颗粒，首先转化为带有荧光染料和胺基的中间体，然后与HIV-Tat肽或鱼精蛋白反应，得到具有膜转位特性的纳米颗粒。我们描述了如何评估最佳的细胞标记与细胞表型和功能的测试。磁荧光纳米颗粒的合成和细胞标记优化可以在48小时内实现，而纳米颗粒的摄取和保留研究通常可能需要长达120小时。标记的细胞可以通过磁共振成像、荧光反射成像、荧光介导的断层扫描、共聚焦显微镜和流式细胞术检测，并且可以基于它们的荧光或磁性特性进行纯化。本方案集中于T细胞标记，但可用于标记各种循环细胞。
Observation of immune and stem cells in their native microenvironments requires the development of imaging agents to allow their in vivo tracking. We describe here the synthesis of magnetofluorescent nanoparticles for cell labeling in vitro and for multimodality imaging of administered cells in vivo. MION-47, a prototype monocrystalline iron oxide nanoparticle, was first converted to an intermediate bearing a fluorochrome and amine groups, then reacted with either HIV-Tat peptide or protamine to yield a nanoparticle with membrane-translocating properties. We describe how to assess optimal cell labeling with tests of cell phenotype and function. Synthesis of magnetofluorescent nanoparticles and cell-labeling optimization can be realized in 48 h, whereas nanoparticle uptakes and retention studies may generally take up to 120 h. Labeled cells can be detected by magnetic resonance imaging, fluorescence reflectance imaging, fluorescence-mediated tomography, confocal microscopy and flow cytometry, and can be purified based on their fluorescent or magnetic properties. The present protocol focuses on T-cell labeling but can be used for labeling a variety of circulating cells.