Imaging the lateral diffusion of membrane molecules with quantum dots
Imaging the lateral diffusion of membrane molecules with quantum dots
复制标题
DOI：
10.1038/nprot.2006.429
复制
发表时间：
2006-01-01
期刊：
影响因子：
14.8
通讯作者：
Triller, Antoine
中科院分区：
文献类型：
--
作者：
Bannai, Hiroko;Levi, Sabine;Triller, Antoine
该协议描述了一种在活细胞中以分子分辨率跟踪膜分子（例如脂质和蛋白质）运动的敏感方法。该技术利用荧光半导体纳米晶体、量子点（QD）作为探针来检测感兴趣的膜分子。量子点的光稳定性和亮度使其能够在单粒子水平上比以前的荧光团（例如荧光蛋白和有机染料）进行更长时间的跟踪。量子点与待跟踪物体的细胞外部分结合，可以用配备光谱灯和敏感冷却电荷耦合器件相机的荧光显微镜记录它们的运动。下面描述的神经元实验过程大约需要 45 分钟。该技术适用于各种培养细胞。
This protocol describes a sensitive approach to tracking the motion of membrane molecules such as lipids and proteins with molecular resolution in live cells. This technique makes use of fluorescent semiconductor nanocrystals, quantum dots (QDs), as a probe to detect membrane molecules of interest. The photostability and brightness of QDs allow them to be tracked at a single particle level for longer periods than previous fluorophores, such as fluorescent proteins and organic dyes. QDs are bound to the extracellular part of the object to be followed, and their movements can be recorded with a fluorescence microscope equipped with a spectral lamp and a sensitive cooled charge-coupled device camera. The experimental procedure described for neurons below takes about 45 min. This technique is applicable to various cultured cells.