An Airyscan 2 confocal laser scanning microscope for the Wolfson Light Microscopy Facility

人类疾病的产生是由于基本生物过程的缺陷。这些过程依赖于细胞内分子之间、组织内细胞之间以及整个生物体中组织之间的相互作用。为了理解和确定有效的治疗方法，我们需要了解生物过程的动态性质，在活细胞中观察这些过程是理解的基础。可视化既验证了其他实验方法，但更重要的是，它经常允许我们提出新的假设。在谢菲尔德大学，生物医学研究人员正在探索分子、细胞和组织的基本行为方式，以了解癌症、感染、炎症、耳聋和神经退行性疾病等疾病中的问题。为了观察生物过程，我们经常将荧光标记附在感兴趣的组分上，并使用荧光显微镜跟踪它们的行为。生物过程的时间尺度是高度可变的：一些过程发生得非常快，这意味着需要快速获取图像，而另一些过程发生在几分钟到几小时的更长时间尺度上。存在与捕获荧光图像相关的许多挑战，因为细胞暴露于光会导致光毒性，这会损害细胞并导致异常行为。此外，一些荧光标记可能非常暗淡，这使得荧光信号难以检测。该提案的目的是为最新的Airyscan 2共聚焦激光扫描显微镜（CLSM）获得资金。这种显微镜不仅将取代现有的两种非常旧的荧光显微镜的功能，而且还将提高我们观察细胞内非常小的结构和/或观察快速发生的高度动态过程的能力。Airyscan 2 CLSM的先进性能将克服目前在理解疾病基础机制方面的局限性。

Human diseases arise because of defects in fundamental biological processes. These processes depend on interactions between molecules within cells, between cells within tissues and tissues in the whole organism. In order to understand and identify effective therapeutics we need to understand the dynamic nature of biological processes and observing such processes in living cells is fundamental to this understanding. Visualisation both validates other experimental approaches but, importantly, often allows us to formulate new hypotheses. At the University of Sheffield biomedical researchers are exploring the fundamental way in which molecules, cells and tissues behave in order to understand what goes wrong in diseases such as cancer, infection, inflammation, deafness and neurodegeneration. In order to observe biological processes we often attach fluorescent labels to components of interest and follow their behaviour using fluorescence microscopes. The time-scale of biological processes is highly variable: some processes happen very quickly, meaning that images need to be acquired speedily while others occur over a much longer time-scale of minutes to hours. There are a number of challenges associated with capturing fluorescence images because the exposure of cells to light can cause phototoxicity which damages cells and causes aberrant behaviour. Additionally, some fluorescent labels can be quite dim which makes the fluorescent signal difficult to detect. The aim of this proposal is to obtain funding for the latest Airyscan 2 confocal laser scanning microscope (CLSM). This microscope will not only replace the functionality of two existing but very old fluorescent microscopes which are difficult to maintain, but will also improve our ability to observe very small structures within cells and/or to observe highly dynamic processes which occur very rapidly. The advanced performance of the Airyscan 2 CLSM will overcome current limitations in understanding fundamental mechanisms that underpin disease.