Multiparametric advanced fluorescence imaging strategies for in situ analysis of live cell signalling

用于活细胞信号传导原位分析的多参数先进荧光成像策略

• 批准号: MR/K015664/1
• 负责人: Malcolm Irving
• 金额: $ 209.96万
• 依托单位: King's College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2013
• 资助国家: 英国
• 起止时间: 2013 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FK015664%2F1
• 关键词: Multiparametric; advanced; fluorescence; imaging; strategies

To understand and combat the causes of human disease, we must understand the basic structure and function of the individual cells that make up the tissues and organs of the human body. For example, to allow the design of effective therapies to target cancer we first need to answer fundamental questions about how the growth, division and movement of cells are controlled. Robert Hooke was the first to use microscopes to describe cell structure in 1665, and since then microscopy has become one of the most powerful tools for cell biologists across the world. The power of light microscopes has of course continued to increase since their invention but, remarkably, the most dramatic improvement has come in the last ten years or so. In that period physicists have worked out how to measure the location of a single protein in a cell with a precision about ten times better that was previously thought possible. This is important because we can now see the internal structure and organisation of cells in much more detail. In parallel, physicists working together with biologists developed microscopical methods that, instead of just producing a map of the locations of one particular protein inside a cell, can produce a map of precisely where protein A is bound to protein B. This is a fundamental advance, because cell function is controlled by pathways and networks of such interactions between specific proteins. Potentially then, these new microscopes provide a window into the internal workings of a cell that allow us to see these protein networks. However, at the moment, the most detailed images can only be obtained from chemically preserved rather than living cells, and each image takes many minutes to record. This is a serious problem, because the interactions between proteins that control cell function take place transiently on the time scale of seconds. To understand cell function, we need movies rather than still images. In the present proposal, biologists and physicists will work together to develop the technology to allow us to record the detailed maps of protein locations and interactions in live cells in milli-seconds rather than minutes or hours. We think that these new developments will unlock the potential of these microscopes to show us how cells work at the molecular level.

为了了解和对抗人类疾病的原因，我们必须了解构成人体组织和器官的单个细胞的基本结构和功能。例如，为了设计针对癌症的有效疗法，我们首先需要回答有关细胞生长、分裂和运动是如何控制的基本问题。罗伯特·胡克在1665年第一个使用显微镜来描述细胞结构，从那时起，显微镜已经成为世界各地细胞生物学家最强大的工具之一。光学显微镜的能力自发明以来当然不断增加，但值得注意的是，最引人注目的改进是在过去十年左右。在此期间，物理学家已经研究出如何测量细胞中单个蛋白质的位置，其精确度比以前认为的可能精度高出约十倍。这很重要，因为我们现在可以更详细地看到细胞的内部结构和组织。与此同时，物理学家与生物学家合作开发了显微镜方法，不仅可以绘制出细胞内特定蛋白质的位置图，还可以绘制出蛋白质A与蛋白质B结合的精确位置图。这是一个根本性的进步，因为细胞功能是由特定蛋白质之间相互作用的途径和网络控制的。因此，这些新的显微镜可能会为我们提供一个了解细胞内部运作的窗口，让我们能够看到这些蛋白质网络。然而，目前，最详细的图像只能从化学保存的细胞而不是活细胞中获得，并且每张图像需要花费许多分钟才能记录下来。这是一个严重的问题，因为控制细胞功能的蛋白质之间的相互作用在几秒钟的时间尺度上短暂发生。要了解细胞功能，我们需要的是电影而不是静止图像。在目前的提议中，生物学家和物理学家将共同努力开发一种技术，使我们能够在毫秒而不是几分钟或几小时内记录活细胞中蛋白质位置和相互作用的详细地图。我们认为，这些新的发展将释放这些显微镜的潜力，向我们展示细胞如何在分子水平上工作。

项目成果

Local dimensionality determines imaging speed in localization microscopy.

• DOI: 10.1038/ncomms13558
• 发表时间: 2017-01-12
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Fox-Roberts P;Marsh R;Pfisterer K;Jayo A;Parsons M;Cox S
• 通讯作者: Cox S

NDP52 activates nuclear myosin VI to enhance RNA polymerase II transcription.

• DOI: 10.1038/s41467-017-02050-w
• 发表时间: 2017-11-30
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Fili N;Hari-Gupta Y;Dos Santos Á;Cook A;Poland S;Ameer-Beg SM;Parsons M;Toseland CP
• 通讯作者: Toseland CP

Recessive mutations in EPG5 cause Vici syndrome, a multisystem disorder with defective autophagy.

• DOI: 10.1038/ng.2497
• 发表时间: 2013-01
• 期刊: NATURE GENETICS
• 影响因子: 30.8
• 作者: Cullup, Thomas;Kho, Ay Lin;Dionisi-Vici, Carlo;Brandmeier, Birgit;Smith, Frances;Urry, Zoe;Simpson, Michael A.;Yau, Shu;Bertini, Enrico;McClelland, Verity;Al-Owain, Mohammed;Koelker, Stefan;Koerner, Christian;Hoffmann, Georg F.;Wijburg, Frits A.;ten Hoedt, Amber E.;Rogers, R. Curtis;Manchester, David;Miyata, Rie;Hayashi, Masaharu;Said, Elizabeth;Soler, Doriette;Kroisel, Peter M.;Windpassinger, Christian;Filloux, Francis M.;Al-Kaabi, Salwa;Hertecant, Jozef;Del Campo, Miguel;Buk, Stefan;Bodi, Istvan;Goebel, Hans-Hilmar;Sewry, Caroline A.;Abbs, Stephen;Mohammed, Shehla;Josifova, Dragana;Gautel, Mathias;Jungbluth, Heinz
• 通讯作者: Jungbluth, Heinz

Fixed pattern noise in localization microscopy.

定位显微镜中的固定图案噪声。

• DOI: 10.1002/cphc.201300756
• 发表时间: 2014
• 期刊: a European journal of chemical physics and physical chemistry
• 作者: Fox-Roberts P

When is an obscurin variant pathogenic? The impact of Arg4344Gln and Arg4444Trp variants on protein-protein interactions and protein stability.

• DOI: 10.1093/hmg/ddab010
• 发表时间: 2021-06-09
• 期刊: Human molecular genetics
• 影响因子: 3.5
• 作者: Fukuzawa A;Koch D;Grover S;Rees M;Gautel M
• 通讯作者: Gautel M