MICA: Imaging of cellular dynamics from single molecules to tissues

显微镜一直是深入了解细胞和组织结构和功能的关键工具。显微镜的使用依赖于对感兴趣的分子/结构进行适当的标记/对比，从而使它们能够被可视化。在过去的20年里，显微镜有了巨大的发展。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”显微镜可以比以往任何时候都能看到更小的结构。这个项目将建立在我们在时间消逝显微镜方面的既定记录的基础上，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专门为显微镜实验而设计的空调房间，以生成细胞的电影。我们使用使萤火虫发光的荧光素酶，当添加化学底物时，使细胞发光(生物发光)。我们还使用了大量不同的荧光蛋白，当它们被不同颜色的光(荧光)照射时，它们会发出不同的颜色。该项目是与领先的显微镜公司卡尔·蔡司的密切合作，卡尔·蔡司将借出设备，一旦设备被优化用于科学应用，大学将购买这些设备。在这个项目中，我们将结合生物发光和荧光，以便可以在单个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启动以制造蛋白质的同时观察细胞信号的计时。我们还将研究一种名为SOFI的新的超分辨率显微镜技术，通过观察单分子发光中的闪烁来定位单分子。这需要数学分析来确定准确的本地化。最后，我们将使用一种名为“SPIM”的显微镜方法来研究三维生物结构。其原理是将一片光照射通过生物样品，然后在与光片平面成90度角的方向上检测到荧光发射。通常，实验室中的细胞是在玻璃上培养的，这显然与人体内的正常情况不同。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。例如炎症、癌症、心血管疾病，以及如何从干细胞生长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。

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.