Ye Dong, Anguissola Sergio, O'Neill Tiina, Dawson Kenneth A
Centre for Bio-Nano Interactions, School of Chemistry and Chemical Biology, University College Dublin, Belfield, Dublin 4, Ireland.
Nanoscale. 2015 Jun 14;7(22):10050-8. doi: 10.1039/c5nr01539a. Epub 2015 May 15.
Subcellular location of nanoparticles has been widely investigated with fluorescence microscopy, via fluorescently labeled antibodies to visualise target antigens in cells. However, fluorescence microscopy, such as confocal or live cell imaging, has generally limited 3D spatial resolution. Conventional electron microscopy can be useful in bridging resolution gap, but still not ideal in resolving subcellular organelle identities. Using the pre-embedding immunogold electron microscopic imaging, we performed accurate examination of the intracellular trafficking and gathered further evidence of transport mechanisms of silica nanoparticles across a human in vitro blood-brain barrier model. Our approach can effectively immunolocalise a variety of intracellular compartments and provide new insights into the uptake and subcellular transport of nanoparticles.
纳米颗粒的亚细胞定位已通过荧光显微镜进行了广泛研究,通过荧光标记抗体来可视化细胞中的靶抗原。然而,荧光显微镜,如共聚焦或活细胞成像,其三维空间分辨率通常有限。传统电子显微镜有助于弥合分辨率差距,但在分辨亚细胞器身份方面仍不理想。利用包埋前免疫金电子显微镜成像,我们对细胞内运输进行了精确检查,并收集了二氧化硅纳米颗粒跨人源体外血脑屏障模型转运机制的进一步证据。我们的方法能够有效地对多种细胞内区室进行免疫定位,并为纳米颗粒的摄取和亚细胞运输提供新的见解。