Spinning disk microscope for Wolfson Light Microscopy Facility

细胞内的成分如何相互作用是细胞行为的关键，这反过来又影响组织的组织，并最终影响整个有机体。观察这些相互作用是如何发生的，对于我们理解健康的有机体是如何形成并继续保持健康的，以及当疾病出现问题时会发生什么，是关键。生物事件的可视化常常使我们提出新的假设。因此，研究人员广泛使用显微镜来了解细胞、组织和生物体的功能。通常，荧光标记被贴在感兴趣的成分上，这样就可以在活细胞中观察到它们。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，而另一些过程则发生在更长的时间尺度上--几分钟到几小时。捕捉荧光图像有许多挑战，因为细胞暴露在光下会引起光毒性，这会损害细胞并导致异常行为。此外，一些荧光标签可能非常模糊，这使得荧光信号很难被检测到。谢菲尔德大学的沃尔夫森光学显微镜设施(LMF)拥有一系列针对特定应用而优化的显微镜。旋转圆盘共聚焦显微镜可以在3个维度上成像快速动态事件。我们目前的旋转圆盘共聚焦显微镜被广泛用于了解正常生理是如何调节的，以及随着生物体的年龄发生了什么变化。它还用于斑马鱼和果蝇等模型生物的实验，帮助我们了解免疫系统是如何工作的。然而，我们现有的系统现在已有12年以上的历史，由于原始部件不再得到制造商的支持，因此很容易受到攻击。这项提议的目的是安装一台替代的旋转圆盘显微镜。这不仅将满足我们现有的需求，而且将开辟一系列令人兴奋的新可能性，因为新一代旋转圆盘共聚焦显微镜在速度、灵敏度和低光毒性方面提供了显著增强的性能。这些优势，再加上捕捉大视野的能力，意味着这是一种非常强大的工具，在LMF中将有广泛的用途。我们已经确定了从细胞和发育生物学、免疫学和微生物学等领域的项目，这些项目将受益于购买最先进的旋转圆盘共聚焦显微镜。

How components interact within cells is key to how cells behave and this in turn affects the organisation of tissues and ultimately the whole organism. Seeing how these interactions occur is key to our understanding of how a healthy organism forms and continues to stay healthy, and what happens when things go wrong in disease. Often visualisation of biological events causes us to formulate new hypotheses. Researchers thus use microscopy extensively to understand how cells, tissues and organisms function. Frequently, fluorescent labels are attached to components of interest which allows them to be observed in living cells. 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 to 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 Wolfson Light Microscopy Facility (LMF) at the University of Sheffield houses a range of microscopes that are optimised for particular applications. Spinning disk confocal microscopy allows imaging of fast dynamic events in 3 dimensions. Our current spinning disk confocal microscope is extensively used to understand how normal physiology is regulated, and what changes as organisms age. It is also used in experiments with model organisms such as zebrafish and fruit flies which help us to understand how the immune system works. Our existing system is however now greater than 12 years old and is vulnerable because the original parts are no longer supported by the manufacturers. The aim of this proposal is to install a replacement spinning disk microscope. This will not only accommodate our existing needs but will open up a range of exciting new possibilities because the new generation spinning disk confocal microscopes offer significantly enhanced performance in terms of speed, sensitivity, and low phototoxicity. These advantages, coupled with the ability to capture wide fields-of-view, mean that this is a very powerful instrument which will have a wide range of uses within the LMF. We have identified projects ranging from cell and developmental biology, immunology and microbiology which will benefit from the acquisition of a state-of-the-art spinning disk confocal microscope.