4D imaging of the dynamic molecular, cellular and tissue organization in living systems

显微镜是揭示生命系统如何组成及其功能的重要工具。最近的进展使研究人员能够以高分辨率看到组织内部和细胞内部。然而，一个限制因素是，所需的强照明会对活样本造成损害，从而减少了对它们的观察时间。一种被称为倒晶格光片显微镜的新技术克服了这个问题，并首次允许对活细胞和组织进行温和的长期成像。这种新型显微镜改变了我们观察活细胞（生命的基本单位）的能力，并检查了它们在健康和疾病中的全部行为。除了低光毒性和高分辨率进入三维多细胞结构，蔡司晶格光片显微镜有一个独特的倒置设计，这是理想的成像细胞和组织培养。这种配置还允许在多孔板中并行跟踪多个样品，从而控制样品偏差，并从单个长期成像实验中生成强大的统计数据集。多井选择可以大大提高实验通量，并实现小规模的化学和遗传筛选。本提案是在埃克塞特大学生物成像中心购买并安装倒置晶格光片显微镜。该显微镜将提供给埃克塞特的生命科学研究人员，以及我们在西南、巴斯、布里斯托尔和卡迪夫的合作大学。使用这种仪器，我们将能够非常详细地看到细胞在发育过程中如何改变身份和构建组织，不同类型的细胞如何交流和合作以提供复杂的功能，以及多细胞组织如何被感染和疾病破坏。埃克塞特生物成像中心有一系列高分辨率显微镜，但缺乏一种能够跟踪多细胞结构或小生物细胞行为的仪器。我们的申请人团队汇集了世界领先的高级研究人员，显微镜技术专家和非常有才华的初级独立调查员。受益于显微镜的尖端研究包括对真菌、植物、海洋无脊椎动物、斑马鱼和人类的研究。研究项目包括分子信号、干细胞命运、胚胎形成、器官发生、药物安全、细胞间通讯、植物病原体相互作用以及纤毛运动的生物物理学。新显微镜获得的大量定量数据将促进生物学家、数学家和物理学家之间的跨学科合作，旨在利用丰富、复杂和令人兴奋的长期成像信息阐明生命规律。

Microscopy is an essential tool for revealing how living systems are composed and how they function. Recent advances have enabled researchers to see deep inside tissues and at high resolution within cells. A limiting factor, however, has been that the powerful illumination required causes damage to living samples, which reduces how long they can be observed. A new technology, called the inverted lattice lightsheet microscope, overcomes this problem and allows for the first time gentle long-term imaging of live cells and tissues. This new microscope is a game changer in our ability to observe living cells, the basic units of life, and examine their full range of behaviours in health and disease. In addition to low phototoxicity and high resolution into 3D multicellular structures, the Zeiss lattice lightsheet 7 microscope has a unique inverted design, which is ideal for imaging cell and tissue cultures. This configuration also allows multiple samples to be tracked in parallel in multiwell plates, which controls for sample bias and generates statistically powerful datasets from a single long-term imaging experiment. The multiwell option can greatly increase experimental throughput and enable small scale chemical and genetic screens.This proposal is to purchase and install an inverted lattice lightsheet microscope in the University of Exeter Bioimaging Center. The microscope will be available to life sciences researchers across Exeter and in our partner Universities in the SouthWest, Bath, Bristol and Cardiff. Using this instrument, we will be able to see in great detail how cells change identity and build tissues during development, how different cell types communicate and cooperate to provide complex functions, and how multicellular tissues are disrupted by infection and disease.The Exeter Bioimaging Centre has a range of high-resolution microscopes but lacks an instrument capable of tracking cell behaviour in multicellular structures or small organisms. Our team of applicants brings together world-leading senior researchers, microscope technology specialists, and highly talented junior independent investigators. The cutting-edge research that will benefit from the microscope includes studies on fungi, plants, marine invertebrates, zebrafish and human. Example projects span investigations into molecular signalling, stem cell fate, embryo formation, organogenesis, drug safety, cell-cell communication, plant pathogen interactions, and the biophysics of ciliary motion. The high volume of quantitative data obtained with the new microscope will foster interdisciplinary collaborations between biologists and mathematicians and physicists aimed at elucidating the rules of life using rich, complex and exciting long-term imaging information.