A confocal microscope for multidisciplinary dynamic studies of complex biological systems

用于复杂生物系统多学科动态研究的共焦显微镜

• 批准号: BB/W019698/1
• 负责人: Conrad Mullineaux
• 金额: $ 49.8万
• 依托单位: Queen Mary University of London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FW019698%2F1
• 关键词: confocal; microscope; multidisciplinary; dynamic; studies

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.

荧光显微镜是探测活细胞和组织组织结构的关键技术。它可用于观察和量化活细胞的动态变化，例如参与细胞分裂、动物胚胎发育或化学信号感知的复杂过程。在过去的几十年中开发的技术通过允许对细胞成分进行特异性标记，如特定的蛋白质，细胞膜或特定的DNA或RNA序列，极大地提高了荧光显微镜的潜力。荧光显微镜的许多变体都是可用的，它们都有自己的优点和缺点。当判断一个特定的荧光显微镜方法对解决一个特定问题的有用性时，关键参数包括空间分辨率（这决定了可以观察到多少细节），时间分辨率（这决定了观察快速过程的能力），光谱分辨率（它决定了可以同时标记和观察多少不同的细胞成分），工作距离（其确定可以成像的样品的厚度）和检测器灵敏度（其确定在样品开始被用于激发荧光的光曝光损坏之前可以观察样品多久）。不同形式的荧光显微镜在这些参数之间具有不同的权衡。共聚焦显微镜是一种多功能的技术，其中通过在1-3个维度上快速扫描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，能够在较厚的样品中分辨出细微的细节，并具有出色的时间分辨率。由于暴露于高强度激光的样品损坏是一个重要的问题，在旧的共焦显微镜，但这个问题大大减少了在最新一代的共焦显微镜使用更高的灵敏度检测器。我们的建议是购买一个非常多功能和国家的最先进的共聚焦显微镜，将提供给用户从整个皇后玛丽，也向外界的学术和工业用户。它将补充其他荧光和电子显微镜技术，使我们能够获得一套技术，帮助我们了解从分子到细胞和组织的生命系统的功能。我们将把它应用于一系列生物学问题，包括细菌细胞的功能，植物对阳光的捕获，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。