Development of the next generation of sicm for live cell imaging

开发用于活细胞成像的下一代 SiCM

• 批准号: BB/D020816/1
• 负责人: David Klenerman
• 金额: $ 37.89万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2007
• 资助国家: 英国
• 起止时间: 2007 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FD020816%2F1
• 关键词: Development; next; generation; sicm; live

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

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响应刺激而变化。目前，我们只能解析冷冻细胞的分子结构，因此无法跟踪动力学，或者我们可以通过使用标记来跟踪活细胞表面上的一种成分。已经开发的一种以足够的分辨率探测活细胞以确定表面上的蛋白质的方法是基于扫描的纳米移液管。这控制在软细胞表面的正上方，使其永远不会接触表面，并可以在表面上光栅化以确定形貌。我们的目标是建立在这种方法的基础上，使其能够更快地对更复杂的细胞表面进行成像，并且还可以在本地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，以检测细胞表面上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。这将使细胞如何在纳米尺度下工作的许多新细节首次被观察到，并开辟了许多新类型的实验

项目成果

A hybrid scanning mode for fast scanning ion conductance microscopy (SICM) imaging.

• DOI: 10.1016/j.ultramic.2012.06.015
• 发表时间: 2012-10
• 期刊: ULTRAMICROSCOPY
• 影响因子: 2.2
• 作者: Zhukov, Alex;Richards, Owen;Ostanin, Victor;Korchev, Yuri;Klenerman, David
• 通讯作者: Klenerman, David

Nanoscale-targeted patch-clamp recordings of functional presynaptic ion channels.

• DOI: 10.1016/j.neuron.2013.07.012
• 发表时间: 2013-09-18
• 期刊: Neuron
• 影响因子: 16.2
• 作者: Novak P;Gorelik J;Vivekananda U;Shevchuk AI;Ermolyuk YS;Bailey RJ;Bushby AJ;Moss GW;Rusakov DA;Klenerman D;Kullmann DM;Volynski KE;Korchev YE
• 通讯作者: Korchev YE

An alternative mechanism of clathrin-coated pit closure revealed by ion conductance microscopy.

• DOI: 10.1083/jcb.201109130
• 发表时间: 2012-05-14
• 期刊: The Journal of cell biology
• 作者: Shevchuk AI;Novak P;Taylor M;Diakonov IA;Ziyadeh-Isleem A;Bitoun M;Guicheney P;Lab MJ;Gorelik J;Merrifield CJ;Klenerman D;Korchev YE
• 通讯作者: Korchev YE

Nanoscale live-cell imaging using hopping probe ion conductance microscopy.

• DOI: 10.1038/nmeth.1306
• 发表时间: 2009-04
• 期刊: NATURE METHODS
• 影响因子: 48
• 作者: Novak, Pavel;Li, Chao;Shevchuk, Andrew I.;Stepanyan, Ruben;Caldwell, Matthew;Hughes, Simon;Smart, Trevor G.;Gorelik, Julia;Ostanin, Victor P.;Lab, Max J.;Moss, Guy W. J.;Frolenkov, Gregory I.;Klenerman, David;Korchev, Yuri E.
• 通讯作者: Korchev, Yuri E.