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

通过晶格光片显微镜实现活细胞超分辨率成像

• 批准号: BB/R000697/1
• 负责人: Franck Pichaud
• 金额: $ 65.39万
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2017
• 资助国家: 英国
• 起止时间: 2017 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FR000697%2F1
• 关键词: Enabling; Live; Cell; Super; Resolution

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.

该应用的目的是建立一个世界领先的活细胞超分辨率成像的卓越极点。细胞是所有复杂生命形式的基本单位，因此了解细胞及其组成部分的生物学是生命科学和医学的关键目标。下一代显微镜正在深刻改变细胞、亚细胞结构和组织的研究方式。比以往任何时候，我们保持领先的研究机构在生物医学科学的能力，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，我们需要深入了解细胞及其成分。实现这一目标的一个主要途径是能够随着时间的推移对大量细胞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进行，以便能够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，光毒性可能导致伪影甚至细胞死亡。先进的光学技术，如光片显微镜，允许在低光毒性的动态研究。在这里，我们要求资金，使我们能够获得和建立一个新的超快速，灵敏，晶格光片显微镜，这将改变我们在细胞和发育生物学的研究能力。该显微镜将为各种各样的研究人员提供解决与正常和疾病条件下的胚胎发育、细胞生物学和神经生物学相关的许多突出问题的机会。为了准确地理解和研究细胞系统，需要在低照度下成像，以最大限度地减少光诱导的样品扰动。光毒性是光学显微镜中的主要问题，已知其诱导伪影和细胞死亡。UCL正在解决这个关键问题，旨在为我们的研究社区提供以高分辨率和最小扰动跟踪细胞的能力。UCL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，UCL与一家小型公司3 i建立了新的令人兴奋的合作伙伴关系，3 i的专业领域是设计和组装最先进的显微镜，包括新一代的光片显微镜，使活细胞成像具有最小的光毒性。UCL和3 i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。在这里，我们请求支持获取LLSM -被认为是同时成像深度，速度，分辨率和标本活力的最佳显微镜。该系统是唯一可通过3 i，并将在UCL细胞和发育生物学的变革，使长期，超分辨率成像的细胞和亚细胞结构和区室。LLSM将补充另一种称为双视图倒置选择性平面照明显微镜（diSPIM）的光片显微镜，该显微镜将由3 i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UCL研究人员设计和实施活体和组织成像的方式。

项目成果

Super-beacons: Open-source probes with spontaneous tuneable blinking compatible with live-cell super-resolution microscopy.

• DOI: 10.1111/tra.12728
• 发表时间: 2020-05
• 期刊: Traffic (Copenhagen, Denmark)
• 作者: Pereira PM;Gustafsson N;Marsh M;Mhlanga MM;Henriques R
• 通讯作者: Henriques R

Quantitative mapping and minimization of super-resolution optical imaging artifacts.

• DOI: 10.1038/nmeth.4605
• 发表时间: 2018-04
• 期刊: Nature methods
• 影响因子: 48
• 作者: Culley S;Albrecht D;Jacobs C;Pereira PM;Leterrier C;Mercer J;Henriques R
• 通讯作者: Henriques R

NanoJ: a high-performance open-source super-resolution microscopy toolbox.

NANOJ：高性能开源超分辨率显微镜工具箱。

• DOI: 10.1088/1361-6463/ab0261
• 发表时间: 2019-04-17
• 期刊: Journal of physics D: Applied physics
• 作者: Laine RF;Tosheva KL;Gustafsson N;Gray RDM;Almada P;Albrecht D;Risa GT;Hurtig F;Lindås AC;Baum B;Mercer J;Leterrier C;Pereira PM;Culley S;Henriques R
• 通讯作者: Henriques R

SRRF: Universal live-cell super-resolution microscopy.

• DOI: 10.1016/j.biocel.2018.05.014
• 发表时间: 2018-08
• 期刊: The international journal of biochemistry & cell biology
• 作者: Culley S;Tosheva KL;Matos Pereira P;Henriques R
• 通讯作者: Henriques R