Institute of Applied Physics and Center for Functional Nanostructures (CFN), Karlsruhe Institute of Technology (KIT), Wolfgang-Gaede-Strasse 1, 76131 Karlsruhe, Germany.
ACS Nano. 2010 Nov 23;4(11):6787-97. doi: 10.1021/nn101277w. Epub 2010 Oct 28.
Uptake and intracellular transport of D-penicillamine coated quantum dots (DPA-QDs) of 4 nm radius by live HeLa cells have been investigated systematically by spinning disk and 4Pi confocal microscopies. Unlike larger nanoparticles, these small DPA-QDs were observed to accumulate at the plasma membrane prior to internalization, and the uptake efficiency scaled nonlinearly with the nanoparticle concentration. Both observations indicate that a critical threshold density has to be exceeded for triggering the internalization process. By using specific inhibitors, we showed that DPA-QDs were predominantly internalized by clathrin-mediated endocytosis and to a smaller extent by macropinocytosis. Clusters of DPA-QDs were found in endosomes, which were actively transported along microtubules toward the perinuclear region. Later on, a significant fraction of endocytosed DPA-QDs were found in lysosomes, while others were actively transported to the cell periphery and exocytosed with a half-life of 21 min.
通过旋转盘和 4Pi 共聚焦显微镜系统地研究了活 HeLa 细胞对 4nm 半径的 D-青霉胺包裹量子点(DPA-QDs)的摄取和细胞内转运。与较大的纳米颗粒不同,这些小的 DPA-QDs 在被内化之前被观察到聚集在质膜上,并且摄取效率与纳米颗粒浓度呈非线性关系。这两个观察结果表明,必须超过一个临界密度阈值才能触发内化过程。通过使用特定的抑制剂,我们表明 DPA-QDs 主要通过网格蛋白介导的内吞作用和较小程度的胞吞作用被内化。DPA-QDs 簇在内涵体中被发现,内涵体沿着微管被主动运输到核周区域。之后,发现内化的 DPA-QDs 的很大一部分在溶酶体中,而其他的则被主动运输到细胞边缘并以 21 分钟的半衰期外排。