MICA: Imaging of cellular dynamics from single molecules to tissues

MICA：从单分子到组织的细胞动力学成像

• 批准号: MR/K015885/1
• 负责人: Michael White
• 金额: $ 162.27万
• 依托单位: University of Manchester
• 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%2FK015885%2F1
• 关键词: MICA; Imaging; cellular; dynamics; single

Microscopy has been a key tool for giving insights into the structure and function of cells and tissues. The use of microscopy depends on achieving appropiate labelling/contrast of the molecules/structures of interest so that they can be visualised. In the past 20 years there have been enormous developments in microscopy. These have included the use of naturally glowing enzymes and proteins to visualise cellular processes. In addition, new "super-resolution" microscopes allow smaller structures to be visualised than ever before.This project will build upon our established track record in timelapse microscopy for the generation of cell movies of biological processes of interest. In the new Systems Microscopy Centre, we have state-of-the-art facilities and air-conditioned rooms specifically designed for microscopy experiments to generate movies of cells. We use the enzyme luciferase that makes fireflies glow to make cells glow (bioluminescence) when a chemical substrate is added. We also use a large number of different fluorescent proteins that glow in different colours when they are illuminated with different coloured light (fluorescence). The project is a close collaboration with the leading microscopy company Carl Zeiss, who will loan equipment which later will be purchased by the University once it has been optimised for the scientific applications.In this project we will combine bioluminescence and fluorescence so that different biological processes can be measured at the same time in single cells. This will allow us to watch cell signal timing at the same time as watching whether and when a gene becomes switched on to make a protein.We will also study a new super-resolution microscopy technique called SOFI where single molecules are located by watching the flickering in their light emission. This requires mathematical analysis to identify the exact localisation. Finally, we will use a microscopy method called "SPIM" that allows 3-dimensional biological structures to be studied. The principle is that a sheet of light is shone through the biological sample and then light emission by fluorescence is detected at 90 degrees to the plane of the light sheet. Often cells in the lab are cultured on glass which is clearly different to what normally happens in the body. Ultimately we hope to improve SPIM and to combine it with bioluminescence and the SOFI super-resolution approach.The project involves a large number of researchers with interests in a range of normal biomedical processes and diseases. Examples include inflammation, cancer, cardiovasular disease, and studies of how to grow new tissues from stem cells and the processes of normal embryo development. This project will act as a focus for future microscopy developments in Manchester University.

显微镜一直是深入了解细胞和组织的结构和功能的关键工具。显微镜的使用取决于对感兴趣的分子/结构进行适当的标记/对比，以便将它们可视化。在过去的 20 年里，显微镜技术取得了巨大的发展。其中包括使用自然发光的酶和蛋白质来可视化细胞过程。此外，新的“超分辨率”显微镜可以比以往更小的结构被可视化。该项目将建立在我们在延时显微镜方面已建立的跟踪记录的基础上，用于生成感兴趣的生物过程的细胞电影。在新的系统显微镜中心，我们拥有最先进的设施和空调室，专门用于显微镜实验以生成细胞电影。当添加化学底物时，我们使用使萤火虫发光的荧光素酶使细胞发光（生物发光）。我们还使用大量不同的荧光蛋白，当用不同颜色的光（荧光）照射时，它们会发出不同颜色的光。该项目是与领先的显微镜公司卡尔蔡司的密切合作，卡尔蔡司将借用设备，一旦这些设备针对科学应用进行了优化，这些设备将由大学购买。在这个项目中，我们将结合生物发光和荧光，以便可以在单细胞中同时测量不同的生物过程。这将使我们能够在观察基因是否以及何时开启以制造蛋白质的同时观察细胞信号时序。我们还将研究一种称为 SOFI 的新超分辨率显微镜技术，其中通过观察光发射的闪烁来定位单个分子。这需要数学分析来确定准确的定位。最后，我们将使用一种称为“SPIM”的显微镜方法来研究 3 维生物结构。其原理是用一片光照射生物样本，然后以与光片平面呈 90 度的方向检测荧光发射的光。实验室中的细胞通常在玻璃上培养，这与体内正常情况明显不同。最终我们希望改进 SPIM 并将其与生物发光和 SOFI 超分辨率方法相结合。该项目涉及大量对一系列正常生物医学过程和疾病感兴趣的研究人员。例子包括炎症、癌症、心血管疾病，以及如何利用干细胞培育新组织和正常胚胎发育过程的研究。该项目将成为曼彻斯特大学未来显微镜发展的重点。

项目成果

Investigating IL-1ß Secretion Using Real-Time Single-Cell Imaging.

使用实时单细胞成像研究 IL-1 分泌。

• DOI: 10.1007/978-1-4939-3566-6_4
• 发表时间: 2016
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Diamond C

Understanding the dynamics of Toll-like Receptor 5 response to flagellin and its regulation by estradiol.

了解Toll样受体5对鞭毛蛋白的反应及其对雌二醇的调节的动力学。

• DOI: 10.1038/srep40981
• 发表时间: 2017-01-23
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Caballero I;Boyd J;Almiñana C;Sánchez-López JA;Basatvat S;Montazeri M;Maslehat Lay N;Elliott S;Spiller DG;White MR;Fazeli A
• 通讯作者: Fazeli A

Integration of Kinase and Calcium Signaling at the Level of Chromatin Underlies Inducible Gene Activation in T Cells.

• DOI: 10.4049/jimmunol.1602033
• 发表时间: 2017-10-15
• 期刊: Journal of immunology (Baltimore, Md. : 1950)
• 作者: Brignall R;Cauchy P;Bevington SL;Gorman B;Pisco AO;Bagnall J;Boddington C;Rowe W;England H;Rich K;Schmidt L;Dyer NP;Travis MA;Ott S;Jackson DA;Cockerill PN;Paszek P
• 通讯作者: Paszek P

Gene-Specific Linear Trends Constrain Transcriptional Variability of the Toll-like Receptor Signaling.

• DOI: 10.1016/j.cels.2020.08.007
• 发表时间: 2020-09-23
• 期刊: Cell systems
• 影响因子: 9.3
• 作者: Bagnall J;Rowe W;Alachkar N;Roberts J;England H;Clark C;Platt M;Jackson DA;Muldoon M;Paszek P
• 通讯作者: Paszek P