Enabling Live-Cell Super Resolution Imaging Through Lattice Light Sheet Microscopy

这个应用程序的目的是建立一个世界领先的极卓越的活细胞超分辨率成像。细胞是所有复杂生命形式的基本单位，因此了解细胞生物学及其组成部分是生命科学和医学的关键目标。下一代显微镜正在深刻地改变研究细胞、亚细胞结构和组织的方式。我们比以往任何时候都更有能力在生物医学科学领域保持领先的研究机构，这要求我们跟上新的、变革性的成像技术。为了进一步了解健康和疾病，我们需要对细胞及其成分有深入的了解。实现这一目标的一个主要途径是能够对大量细胞随时间变化的行为和功能特性进行成像。这必须在高空间和时间分辨率下完成，以便发现和研究动态过程。重要的是，细胞和组织的成像也必须在防止光毒性的同时进行，这可能导致伪影甚至细胞死亡。先进的光学技术，如薄片显微镜，允许在低光毒性下进行这种动态研究。在此，我们请求资金，使我们能够获得并建立一种新型的超快速、超灵敏的点阵光片显微镜，这将改变我们在细胞和发育生物学方面的研究能力。该显微镜将被广泛的研究人员使用，以解决与正常和疾病条件下胚胎发育、细胞生物学和神经生物学有关的许多悬而未决的问题。为了准确地理解和研究细胞系统，需要在低照度下成像，最大限度地减少样品的光致扰动。光毒性是光学显微镜中的一个主要问题，它会引起伪影和细胞死亡。伦敦大学学院正在解决这个关键问题，目的是为我们的研究界提供高分辨率和最小扰动的细胞跟踪能力。伦敦大学学院在开发和实施解决光毒性问题的光学方法方面发挥了重要作用。近年来，伦敦大学学院与一家小型公司3i建立了令人兴奋的合作伙伴关系，该公司的专业领域是设计和组装最先进的显微镜，包括新的产生光片显微镜，使活细胞成像具有最小的光毒性。UCL和3i共同参与联合项目，旨在开发通过光片显微镜（LLSM）实现超分辨率长期成像的新技术。在此，我们请求获得LLSM的支持，LLSM被认为是同时成像深度、速度、分辨率和标本存活率最好的显微镜。该系统是通过3i唯一可用的，通过实现细胞、亚细胞结构和隔室的长期、超分辨率成像，将对UCL的细胞和发育生物学产生革命性的影响。LLSM将补充另一种称为双视图倒置选择平面照明显微镜（diSPIM）的光片显微镜，该显微镜将由3i在LMCB组装，作为我们合作协议的一部分。这种特殊的仪器组合将改变伦敦大学学院研究人员设计和实施活体和组织成像的方式。

The aim with this application is to set up a world-leading pole of excellence for live-cell Super-Resolution imaging. The cell is the fundamental unit of all complex life forms and so understanding the biology of the cell and its component parts is a key goal in life science and medicine. Next-generation microscopes are profoundly changing the way cells, sub-cellular structures, and tissues can be studied. More than ever before, our ability to remain a leading research institution in biomedical sciences requires us to stay abreast of new, transformative imaging technologies. To further our understanding of health and diseases requires a deep knowledge of the cell and its constituents. One main path toward this goal is to be able to image the behavior and functional properties of large populations of cells over time. This must be done at high spatial and temporal resolution so that dynamic processes can be discovered and studied. Importantly, imaging of cells and tissues must also be done while preventing photo-toxicity, which can lead to artefacts and even cell death. Advanced optical techniques such as light sheet microscopy, allow for such dynamic studies at low photo-toxicity. Here we request funds to enable us to acquire and set up a novel ultra-fast, exquisitely sensitive, lattice light sheet microscope that will transform our research capabilities in cell and developmental biology. The microscope will be accessible to a wide variety of researchers addressing many outstanding questions relating to embryonic development, cell biology and neurobiology in both normal and disease conditions.To accurately understand and study cellular systems requires imaging at low-illumination minimizing light-induced perturbation of the sample. Photo-toxicity is a main issue in light microscopy, that is know to induce artefacts and cell death. This critical problem is being addressed at UCL with the aim to provide our research-community with the capacity to follow cells at high-resolution and with minimal perturbation. UCL has been instrumental in developing and implementing optical approaches tackling the problem of photo-toxicity. In the recent years, UCL has entered a new and exciting partnership with a small size company, 3i, whose area of expertise is to design and assemble state-of-the-art microscope, including new generating light sheet microscopes which enable live cell imaging with minimum photo-toxicity. Together UCL and 3i are engaged in joint projects that aim at developing new technologies enabling Super-Resolution long-term imaging, through light-sheet microscopy (LLSM). Here we request for support to acquire a LLSM - considered the best performing microscope for simultaneous imaging depth, speed, resolution and specimen viability. This system is uniquely available through 3i and will be transformative for cell and developmental biology at UCL by enabling long-term, Super-Resolution imaging of cells and subcellular structures and compartment. The LLSM will complement another type of light sheet microscope called Dual-View Inverted Selective Plane Illumination Microscope (diSPIM), which will be assembled by 3i at the LMCB as part of our collaborative agreement. This particular combination of instruments will transform the ways UCL researchers are designing and implementing live and tissue imaging.