A Single Molecule Detection Platform (SMD) for a Leica SP8 TCS to analyse protein-protein interactions in living specimen.

从某些水母中发现的荧光蛋白GFP(绿色荧光蛋白)，彻底改变了我们研究蛋白质定位、蛋白质动力学和蛋白质相互作用的能力。这些和相关的荧光蛋白使我们能够研究活细胞复杂环境中的蛋白质功能。在荧光蛋白生物学发展的同时，荧光成像方法和显微系统也取得了进展，使得定位与这些荧光蛋白相连的蛋白质，定量它们的丰度，并探索它们的流动性和相互作用成为可能。最近，两种成像方法，如Förster共振能量转移(FRET)和荧光相关光谱(FCS)进行了改进，以便可以在用户友好的商业激光扫描显微镜上完成，取代了对复杂定制显微镜的需求。绿色荧光蛋白生物学和成像方法的共同进步为研究活细胞中蛋白质的动态性质提供了巨大的刺激。埃克塞特大学在生物医学研究的许多方面都表现出色，从真菌相关的植物疾病研究到脊椎动物胚胎发育的信号生物学。在过去，生化方法被用来研究蛋白质之间的相互作用。这些观察结果现在可以在活细胞中实时补充和扩展。因此，最新一代的单分子探测器改变了蛋白质生物学--细菌、真菌、植物和动物--的游戏规则。现在，使用荧光显微镜可以生成活细胞中蛋白质相互作用的动态地图。在这一应用中，我们寻求支持购买一个现代化的单分子检测平台(SMD)，该平台将补充埃克塞特生物成像中心的现有设施。

The discovery of the fluorescent protein GFP (green fluorescent protein) from certain jellyfish, has revolutionized our ability to study protein localization, protein dynamics and interactions of proteins. These and related fluorescent proteins allow us to investigate protein function within the complex environment of the living cell. Parallel to the developments in fluorescent protein biology, there have been advances in fluorescence imaging methods and microscopical systems that make it possible to localize proteins linked with such fluorescent proteins, to quantitate their abundance and to probe their mobility and interactions with each other. Recently, two imaging methods such as Förster resonance energy transfer (FRET) and fluorescence correlation spectroscopy (FCS) have been modified so that they can be done on user-friendly, commercially available laser scanning microscopes, replacing the need for complex custom-built microscopes. The combined advances in GFP biology and imaging methods are providing a massive stimulus for investigating the dynamic properties of proteins in living cells. The University of Exeter excels in many aspects of biomedical research, from fungal-related plant disease research to signalling biology in vertebrate embryonic development. In the past, biochemical approaches have been used to investigate protein-protein interactions. These observations can now be complemented and extended in real time in living cells. The latest generation of single molecule detectors is therefore a game changer for protein biology - in bacteria, fungi, plants and animals. It is now possible to generate dynamic maps of protein interactions in living cells using a fluorescence microscope. In this application, we seek support to purchase a modern single molecule detection platform (SMD), which will complement the existing facilities at the Exeter Bioimaging Centre.