Development of single-molecule tracking using fluorescent silicon nanoparticles and its application to signaling mechanisms in clathrin-coated pits

细胞膜上的单个受体分子被荧光硅纳米颗粒特异性标记，没有眨眼或光漂白，这使得首次有可能通过笼罩蛋白包裹的凹坑来可视化个体的内化事件。建立了一种将酶标记物的配体偶联到纳米颗粒表面的方法，从而提高了探针的通用性。一种新的摄像系统被进一步开发，使我们能够在0。1毫秒分辨率。

Single receptor molecules in the plasma membrane were specifically labeled with fluorescent silicon nanoparticles without blinking or photobleaching, making it possible for the first time to visualize individual internalization events through clathrin-coated pits. A method for conjugating ligands for enzyme-based protein tag to nanoparticle surface was developed, so that the versatility of the probe was enhanced. A new camera system was further developed, allowing us to detect each elementary process of receptor association and signaling complex formation at a 0. 1-ms resolution.