High-throughput Lattice Light Sheet Microscopy : Imaging Across Scales.

生物医学研究人员经常依靠现场直播电影来了解疾病的亚细胞原因，并开发任何治疗干预措施。这种记录活细胞和胚胎的电影可能会对样本产生高度的光毒性，并导致细胞在几个小时或一天内死亡。为了克服这一障碍，研究人员已经开始使用格子光片显微镜(LLSM)，这种显微镜的照明方法极其温和和快速。在大伦敦地区，我们目前在任何多用户设施中都没有能够进行长达几天或一周的高分辨率成像的晶格光片显微镜。这是一个巨大的技术劣势，因为有几个集团正在求助于短期隔夜电影，而这些可以简单地扩展到长期电影，而不需要额外的人员或消耗品费用投资。我们的目标是建立一个多用户LLSM，用于长期和高分辨率的活细胞研究。在MRC为各种模式生物(从细菌到人类细胞)的研究提供资金的环境中，这是一个及时且独特的经济高效的解决方案，可以在英国实现世界级的生物医学研究和培训。十名在MRC研究领域学习的生物医学研究人员(由MRC资助)旨在通过利用蔡司LLS7这一配备双摄像头的高通量显微镜来扩展他们正在进行的研究。首先，有了LLS7，该联盟将能够摆脱短期电影，开始录制长期电影，这样任何亚细胞损伤的原因和后果都可以在2D和3D细胞培养中精确记录下来。其次，利用LLS7，研究人员将开始跟踪蠕虫、苍蝇和鱼类胚胎早期阶段的胚胎进化，以精确地将细胞与细胞之间的相互作用与细胞命运决定联系起来。第三，MRC资助的结构生物学家还计划进行高通量的结构-功能研究，以加快评估突变和药物治疗条件在细胞中的即时和长期影响的进程。这三个目标将共同推进分子、细胞、发育和结构生物学领域正在进行的生物医学研究。此外，通过与人工智能专家(大学和行业)的合作，该联盟将在高通量单细胞图像组学这一新兴领域为新的自动化数据分析工具铺平道路。考虑到长期成像系统将在正在进行的研究中产生的积极影响，QMUL已同意为该设备提供全额匹配资金。除了加速正在进行的研究，所要求的LLS7显微镜还支持玛丽女王在发展数百万精密医疗研究所方面的雄心勃勃的努力。当该研究所的单细胞生物学中心准备就绪时，LLS7的用户可以扩展他们的工作，开始使用动态标记来研究患者细胞的命运。当这个框架可用时，可以帮助彻底改变治疗策略--这是一个前沿的医疗领域。作为生物医学研究的高通量实时成像领域的领导者，可以帮助发展这一长期努力，与英国国民健康保险制度(例如，新生儿或未来的医疗团队)建立高度有影响力的合作(S)。具体地说，与临床医生合作，该设备可用于以下用途：(A)设计单细胞诊断管道，以观察从血液、口腔拭子或尿液中提取的细胞的命运；(B)使用呈现疾病相关变异的患者来源细胞建立细胞疾病模型；以及(C)使用基于单细胞的表型逆转分析，诊断此类疾病相关变异的影响，以实现精确医疗保健。总而言之，确保目前在欧洲无法获得的LLS7，将加强正在进行的国际领先的生物医学研究，并从长远来看，为建立面向患者的诊断战略提供一条途径。

Biomedical researchers frequently rely on live movies to understand the subcellular cause of diseases and to develop any therapeutic interventions. Such recording of movies of live cells and embryos can be highly phototoxic to the sample and result in cell death within a few hours or a day. To overcome this hurdle, researchers have started using lattice light-sheet microscopes (LLSM) that are extremely gentle and extremely fast with their illumination methodology. In the Greater London Region, we currently do not have a lattice light-sheet microscope in any multi-user facility which is capable of long-term and high-resolution imaging for up to a few days or a week. This is a huge technical disadvantage as several groups are resorting to short-term overnight movies, while these could simply be extended into long-term movies without additional investment into personnel or consumables expenses.We aim to establish a multi-user LLSM for long-term and high-resolution live-cell studies. In an environment where MRC funds research in a variety of model organisms (from bacteria to human cells), this is a timely and uniquely cost-effective solution to enable World-class Biomedical research and training in the UK.Ten biomedical researchers studying in a variety of MRC research areas (and funded by MRC) aim to extend their ongoing studies by taking advantage of the Zeiss LLS7, a dual camera-equipped high-throughput microscope. First, with an LLS7, the consortium will be able to move away from short-term movies and start recording long-term movies so that the cause and consequence of any subcellular lesion can be precisely documented in 2D and 3D cell cultures. Second, using the LLS7, the researchers will start tracking the evolution of embryos in the early stages of the worm, fly and fish embryos to precisely link cell-cell interactions and cell fate decisions. Third, MRC-funded structural biologists are also planning to perform high-throughput structure-function studies to accelerate the process of evaluating the immediate and long-term impact of mutants and drug treatment conditions in cells. Together, the three objectives will advance ongoing biomedical research in molecular, cell, developmental and structural biology fields. In addition, by collaborating with Artificial Intelligence experts (in the university and industry) the consortium will set the path for new automated data analysis tools in the emerging area of high-throughput single-cell 'image-omics' field.Considering the positive impact a long-term imaging system would make in ongoing research, QMUL has agreed to fully match fund the equipment. In addition to accelerating ongoing research, the requested LLS7 microscope supports Queen Mary's ambitious effort in developing a multimillion Precision Healthcare Institute. When the single-cell biology hub of the institute is ready, LLS7 users could extend their work to start studying patient cell fate using dynamic markers. This framework, when available, can help revolutionise therapy strategies - a frontiers healthcare area. Being a leader in high-throughput live imaging for biomedical research could help develop this long-term effort in building highly impactful collaboration(s) with the NHS (for example, newborn or future health teams). Specifically, in collaboration with clinicians, the equipment could be used for the following: (a) to design a single-cell diagnostics pipeline to observe the fate of cells recovered from blood, buccal swabs or urine, (b) to build cellular disease models using patient-derived cells presenting disease-oriented variants and (c) to diagnose the impact of such disease-oriented variants using single-cell based phenotypic reversal assays for precision healthcare. In summary, securing the LLS7 which is currently unavailable in Europe, will strengthen ongoing internationally leading biomedical research, and in the long-term provide a path to building a patient-facing diagnostic strategy.