A Bessel beam light sheet microscope

贝塞尔光束光片显微镜

• 批准号: BB/S019065/1
• 负责人: Susan Cox
• 金额: $ 44.39万
• 依托单位: King's College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FS019065%2F1
• 关键词: Bessel; beam; light; sheet; microscope

Fluorescence microscopy has become a very important technique in the biosciences because it allows the distributions of specific proteins to be imaged. The resolution it provides is not as good as electron microscopy, but it has the major advantage that it can be used in live cells. However, the imaging process can still have a toxic effect on cells, and this is a major limitation in cell biology studies.A popular way to minimise the effects of imaging on cells is called light sheet microscopy. This only illuminates the part of the sample that is in focus, meaning many more pictures can be taken without damaging the cell. Generally, this also makes the resolution worse - but an approach has recently been developed which not only illuminates using a light sheet, but also improves the resolution. This is achieved using Bessel beams, which use interference and are therefore not subject to the same limitations as most microscopy techniques. Using this lattice light sheet approach, the sample can be imaged quickly and with excellent resolution.We will build a replica of the lattice light sheet system at Janelia Research Campus (where the technique was developed) in the Microscopy Innovation Centre (MIC) at King's College London, using blueprints and equipment lists provided to us by Janelia. The MIC has been created by KCL in recognition of the central role that microscopy plays in biological and biomedical innovation. It's purpose is to make advanced microscopy techniques available to users, and this is made possible by two experienced optical microscopy specialists. They will build the system, maintain it, and make it available to users. A particularly notable feature of this system is that it will have an extra module to allow specific areas of the samples to either be bleached or for fluorophores which can emit in two different colours to be switched from one to the other. We believe this would be the first system of its kind in the UK.A wide range of different project applications have been proposed. These include three from groups who already travel to Janelia Research Campus to use the system there, so they are familiar with the equipment and sample preparation requirements. Projects will improve understanding of various basic biological processes: cell migration, cell division, how cancer cells invade, the characteristics of neurons which have been derived from stem cells, and the cytoskeletal structure and mitochondria in heart cells. The lattice light sheet microscope will let us image these live systems at very high resolution, and observe how they change over time, enabling new biological discoveries. After an initial evaluation period where the microscope is only available to users at King's, the microscope will be opened to users in other institutions, who would them be able to access this system with full technical support without travelling to the USA. The ability to image cells at high resolution, without the the imaging process changing their behaviour, promises to vastly increase the amount of information available about cells. By making so much more information available, this microscope will help to maintain the UK's leading position in cell biology and biomedical research.

荧光显微镜已经成为生物科学中非常重要的技术，因为它允许对特定蛋白质的分布进行成像。它提供的分辨率不如电子显微镜，但它的主要优点是可以用于活细胞。然而，成像过程仍然会对细胞产生毒性作用，这是细胞生物学研究的一个主要限制。一种最大限度地减少成像对细胞影响的流行方法称为光片显微镜。这只会照亮聚焦的样品部分，这意味着可以在不损坏细胞的情况下拍摄更多照片。通常，这也使分辨率变差，但是最近已经开发出一种方法，该方法不仅使用光片照明，而且还提高了分辨率。这是使用贝塞尔光束实现的，贝塞尔光束使用干涉，因此不受大多数显微镜技术相同的限制。我们将在伦敦国王学院的显微镜创新中心（MIC）的Janelia研究园区（该技术是在那里开发的）建立一个晶格光片系统的复制品，使用Janelia提供给我们的蓝图和设备清单。MIC由KCL创建，以表彰显微镜在生物和生物医学创新中发挥的核心作用。它的目的是使先进的显微技术提供给用户，这是由两个经验丰富的光学显微镜专家。他们将构建系统，维护它，并将其提供给用户。该系统的一个特别值得注意的特点是，它将有一个额外的模块，允许样品的特定区域被漂白，或者允许可以发射两种不同颜色的荧光团从一种颜色切换到另一种颜色。我们相信这将是英国第一个此类系统。已经提出了广泛的不同项目应用。其中包括三个来自已经前往Janelia研究园区使用该系统的团体，因此他们熟悉设备和样品制备要求。项目将提高对各种基本生物过程的理解：细胞迁移，细胞分裂，癌细胞如何入侵，干细胞衍生的神经元的特征，以及心脏细胞中的细胞骨架结构和线粒体。晶格光片显微镜将让我们以非常高的分辨率对这些活系统进行成像，并观察它们如何随时间变化，从而实现新的生物学发现。经过初步评估期（该显微镜仅向King's的用户开放）后，该显微镜将向其他机构的用户开放，他们无需前往美国即可访问该系统并获得全面的技术支持。在成像过程不改变细胞行为的情况下，以高分辨率成像细胞的能力有望大大增加有关细胞的可用信息量。通过提供更多的信息，这种显微镜将有助于保持英国在细胞生物学和生物医学研究方面的领先地位。