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，这种排列随细胞执行特定功能和对刺激作出反应的时间而变化。目前我们只能在冷冻细胞上解析分子结构，因此不能跟踪动态，或者我们只能通过标记来跟踪活细胞表面的一个成分。一种以足够的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吸管。这控制在柔软的细胞表面上方，这样它就不会接触到表面，并且可以在表面上光栅来确定地形。我们的目标是在这种方法的基础上，使其能够更快地成像更复杂的细胞表面，并且还可以在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来检测细胞表面的单个蛋白质复合物，它们的组织以及它们如何随时间变化。这将使人们第一次观察到细胞在纳米尺度上如何工作的许多新细节，并开启许多新型实验

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