The University of Bath Multi-user Light Sheet Fluorescence Microscopy

我们希望为巴斯大学的显微镜设施增加一个重要的尖端仪器。生理学、细胞和发育生物学的许多方面都是动态的，包括秒（例如心跳）、分钟（例如细胞迁移）和小时甚至天的时间尺度（例如胚胎发育和植物生长）。同样，生物系统通常是三维的，其结构安排与其功能（例如心血管系统，大脑，胚胎）不可分割。因此，为了详细研究生物过程，我们需要显微镜，使我们能够在这个时间尺度范围内对生物过程进行成像，并在完整的3D样品（整个生物体或组织）中进行成像。一种新的技术，光片荧光显微镜（LSFM），允许所有这些，并承诺彻底改变这些领域。LSFM设计的关键方面确保快速成像，以便可以解决秒-分钟时间尺度上的变化;温和成像，以便标本在几分钟，几小时甚至几天的长期研究中不会损坏;更深的成像，以便更厚的标本和整个生物体变得可访问。这意味着我们可以研究在长时间内对药物或病原体的反应变化;我们可以监测细胞在复杂组织甚至整个生物体中的迁移，检测它们在环境改变时的行为变化;我们可以研究活细胞中的过程，这些过程被组织成三维结构，更准确地模拟生理环境。在巴斯，我们建议在七个初始项目中使用LSFM，每个项目都使用LSFM的一个或多个关键功能：研究干细胞在斑马鱼胚胎发育中的作用，脂肪发育对小鼠模型体重的控制，细胞环境对软骨特性的影响，细菌群落如何（生物膜）在逃避抗生素处理方面变得更有效，植物根对感染的反应，如何制造可再生的类器官培养物，以及绘制小鼠大脑中的神经元活动。该仪器将被集成到巴斯大学的显微镜和分析套件中，以确保专家用户培训和设备支持。它将支持巴斯大学和我们的合作机构/组织内广泛的跨学科研究社区。

We wish to add an important cutting-edge instrument to the University of Bath's microscopy facilities. Many aspects of physiology, cell and developmental biology are dynamic, including those on second (e.g. heart-beat), minute (e.g. cell migration) and hour or even day timescales (e.g. embryonic development and plant growth). Likewise, biological systems are frequently 3-dimensional, with structural arrangements being integral to their function (e.g. cardiovascular systems, brain, embryos). Thus, to study biological processes in detail, we need microscopy that can allow us to image biological processes on this range of timescales, and in intact 3-D samples (whole organisms, or tissues). A new technique, Light Sheet Fluorescent Microscopy (LSFM), allows all of these and promises to revolutionise these areas. Key aspects of the LSFM design ensure fast imaging, so that changes on time-scales of seconds-minutes can be resolved; gentle imaging, so that specimens are not damaged over long-term study for minutes, hours or even days; and deeper imaging, so that thicker specimens and whole living organisms become accessible. It means that we can study changes in response to drugs or pathogens over long timescales; that we can monitor cells as they migrate through complex tissues or even whole organisms, detecting changes in their behaviour as their environment alters; and that we can study processes in living cells organised into 3-D structures that more accurately mimic a physiological environment. In Bath, we propose using the LSFM for seven initial projects, each utilising one or more key features of LSFM: to study stem cells in developing zebrafish embryos, fat development in mouse models of body mass control, cell environment on cartilage properties, how bacterial communities (biofilms) become more efficient at evading antibiotic treatment, plant root responses to infection, how to make reproducible organoid cultures, and mapping neuronal activation in mouse brains. The instrument will be integrated into the Microscopy and Analysis Suite in the University of Bath, ensuring expert user training and equipment support. It will support a broad, interdisciplinary research community within the University of Bath and our collaborator institutions/organisations.