A confocal microscope for multidisciplinary dynamic studies of complex biological systems

荧光显微镜是探测活细胞和组织结构的关键技术。它可以用来观察和量化活细胞的动态变化，例如参与细胞分裂、动物胚胎发育或感知化学信号的复杂过程。过去几十年发展起来的技术极大地提高了荧光显微镜的潜力，因为它允许对细胞成分进行特定标记，如特定的蛋白质、细胞膜或特定的DNA或RNA序列。荧光显微镜有许多变种，它们都有各自的优点和缺点。在判断一种特定的荧光显微镜方法对于解决特定问题的有用性时，关键参数包括空间分辨率(决定可以观察到多少细微细节)、时间分辨率(决定观察实际快速过程的能力)、光谱分辨率(决定可以同时标记和观察多少不同的细胞组件)、工作距离(确定样品可以成像的厚度)和探测器灵敏度(决定样品在受到用于激发荧光的光照射之前可以被观察到多长时间)。不同形式的荧光显微镜在这些参数之间有不同的权衡。共焦显微镜是一种多功能技术，它通过快速扫描1-3维的高度聚焦的激光光斑来对样品进行成像。它缺乏一些最近开发的荧光显微镜形式的极高空间分辨率，但它提供了特别好的光谱分辨率，能够在较厚的样本中解析精细细节和出色的时间分辨率。暴露在高强度激光下的样品损坏在老式共焦显微镜中是一个重要的问题，但在最新一代的共焦显微镜中，通过使用更高灵敏度的探测器，这个问题大大减少了。我们的建议是购买一台多功能的、最先进的共焦显微镜，它将向玛丽女王各地的用户以及外部学术和工业用户提供。它将补充这里提供的其他荧光和电子显微镜技术，使我们能够使用一套技术，帮助我们了解从分子到细胞和组织的各种规模的生命系统的功能。我们将把它应用于广泛的生物学问题，包括细菌细胞的功能、植物对阳光的捕捉、细胞对化学信号的感知、细胞分裂时染色体的重组以及动物胚胎的发育。

Fluorescence microscopy is a key technique for probing the organisation of living cells and tissues. It can be used to observe and quantify dynamic changes in live cells, for example the complex processes in involved in cell division, the development of animal embryos or the perception of chemical signals. Techniques developed over the last decades have hugely increased the potential of fluorescence microscopy by allowing the specific labelling of cell components such a specific proteins, cell membranes or particular DNA or RNA sequences. Many variants of fluorescence microscopy are available, all with their own advantages and drawbacks. The crucial parameters when judging the usefulness of a particular fluorescence microscopic method for tackling a particular problem include the spatial resolution (which determines how much fine detail can be observed), the time resolution (which determines the ability to observe rapid processes in action), the spectral resolution (which determines how many distinct cell components can be labelled and observed simultaneously), the working distance (which the determines the thickness of the sample that can be imaged), and the detector sensitivity (which determines how long a sample can be observed before it starts to be damaged by exposure the light used to excite fluorescence). Different forms of fluorescence microscopy have different trade-offs between these parameters. Confocal microscopy is a versatile technique in which the sample is imaged by rapidly scanning a highly-focused laser spot across it, in 1-3 dimensions. It lacks the extremely high spatial resolution of some recently-developed forms of fluorescence microscopy, but it provides particularly good spectral resolution with an ability to resolve fine details within thicker samples and excellent time resolution. Sample damage due to exposure to high-intensity lasers was a significant issue in older confocal microscopes, but this problem is greatly decreased in the latest generation of confocal microscopes by the use of higher-sensitivity detectors. Our proposal is to purchase a very versatile and state-of-the-art confocal microscope that will be made available to users from across Queen Mary and also to outside academic and industrial users. It will complement the other fluorescence and electron microscopic techniques available here to give us access to a suite of techniques that will help us to understand the function of living systems on scales from molecules up to cells and tissues. We will apply it to a huge range of biological problems, including the function of bacterial cells, the trapping of sunlight by plants, the perception of chemical signals by cells, the re-organisation of chromosomes as a cell divides and the development of animal embryos.