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

生物医学研究人员经常依靠现场电影来了解疾病的亚细胞原因并开发任何治疗干预措施。活细胞和胚胎的这种电影记录可能对样品具有高度的光毒性，并导致细胞在几小时或一天内死亡。为了克服这一障碍，研究人员已经开始使用点阵光片显微镜（LLSM），这种显微镜的照明方法非常温和，速度非常快。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长期和高分辨率成像的点阵光片显微镜。这是一个巨大的技术劣势，因为有几个团队采用短期过夜电影，而这些电影可以简单地扩展到长期电影，而无需额外的人员或耗材费用投资。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资助各种模式生物研究的环境中，（从细菌到人体细胞），这是一个及时和独特的成本效益的解决方案，使世界一流的生物医学研究和培训在英国。十个生物医学研究人员在各种MRC研究领域的研究（并由MRC资助）旨在通过利用蔡司LLS 7（一种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LLS 7，该联盟将能够摆脱短期电影，开始记录长期电影，以便在2D和3D细胞培养中精确记录任何亚细胞病变的原因和后果。其次，使用LLS 7，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，以精确地将细胞间相互作用和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结构-功能研究，以加速评估突变体和药物治疗条件对细胞的直接和长期影响的过程。这三个目标将共同推动分子、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（大学和工业界）的合作，该联盟将为高通量单细胞“图像组学”领域的新兴领域新的自动化数据分析工具开辟道路。考虑到长期成像系统将对正在进行的研究产生积极影响，QMUL已同意为设备提供全额匹配资金。除了加速正在进行的研究，LLS 7显微镜还支持玛丽女王雄心勃勃的努力，建立一个数百万人的精密医疗保健研究所。当该研究所的单细胞生物学中心准备就绪时，LLS 7用户可以扩展他们的工作，开始使用动态标记物研究患者细胞的命运。这个框架，当可用时，可以帮助革命性的治疗策略-一个前沿的医疗保健领域。作为生物医学研究的高通量实时成像的领导者，可以帮助开发这种长期的努力，与NHS（例如，新生儿或未来的健康团队）建立高度影响力的合作。具体而言，与临床医生合作，该设备可用于以下方面：（a）设计单细胞诊断管道，以观察从血液、口腔拭子或尿液中回收的细胞的命运，（B）使用呈递疾病导向变体的患者来源的细胞构建细胞疾病模型，和（c）使用单克隆抗体诊断这种疾病导向变体的影响。基于细胞的表型逆转试验，用于精准医疗保健。总之，确保LLS 7目前在欧洲不可用，将加强正在进行的国际领先的生物医学研究，并在长期内提供一条建立面向患者的诊断策略的途径。

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.