Development of the next generation of sicm for live cell imaging

细胞表面是许多分子的复杂排列，这种排列随着细胞执行特定功能和对刺激的反应而变化。目前，我们只能对冷冻细胞的分子结构进行解析，因此不能跟踪动力学，或者通过标记只能跟踪活细胞表面的一个组分。一种已经开发出来的以足够分辨率探测活细胞以确定表面蛋白质的方法是基于扫描的纳米管。这就控制在柔软的细胞表面上，这样它就永远不会接触，可以在表面上栅格来确定地形。我们的目标是建立在这种方法的基础上，这样它就可以更快地对更复杂的细胞表面进行成像，并可以局部应用试剂来识别我们检测到的特征。这将以足够的分辨率完成，以检测细胞表面的个别蛋白质复合体，它们的组织以及这种复合体如何随时间变化。这将允许首次观察到细胞如何在纳米尺度上工作的许多新细节，并打开许多新类型的实验

The cell surface is a complex arrangement of many molecules and this arrangement changes in time for the cell to perform specific functions and in response to stimuli. At present we can only resolve the molecular structure on frozen cells, so dynamics cannot be followed or we can follow just one component on the surface of live cells by the use of labelling. One method that has been developed to probe live cells at sufficient resolution to determine proteins on the surface is based on a scanned nanopipette. This controlled just above the soft cell surface so that it never touches and can be rastered over the surface to determine the topography. We aim to build on this method so that it can image more complex cell surfaces faster and can also locally apply reagents to identfy the features that we detect. This will be done with sufficient resolution to detect indvidual protein complexes on the cell surface, their organization and how this changes with time. This will allow many new details of how the cell works at the nanoscale to be observed for the first time and opening up many new types of experiments