Light-sheet microscopy for 3D bioimaging of engineered tissues and microphysiological systems

近年来，随着生物制造和组织工程技术的快速发展，我们越来越多地被要求可视化复杂的工程组织和3D生物外植体，以满足不断扩大的生物学研究前沿。然而，传统的共聚焦显微镜在扫描大的3D组织样本时速度极慢，无法实时跟踪实时的3D生物过程。这些限制可以通过光片显微镜来克服，光片显微镜用一片光照射组织样本，以确保信号只从聚焦区域发出。这种机制减少了样品的总曝光量，并将扫描速度提高了数个数量级，使复杂组织的长期实时成像成为可能。虽然我们的联合申请者小组在组织工程、芯片器官、生物打印、微生物建模和药物输送方面的发展取得了重大进展，但我们现在严重限制了我们以3D方式表征和研究组织样本的能力。因此，所要求的光片显微镜对于探索关键的特征问题和实现新的生物学研究是必不可少的。*由于这项新兴技术的稀有，在组织工程和芯片上的器官中使用光片显微镜仍处于初级阶段。为了将这项技术应用到我们的协作实验室空间，光片系统应该是模块化的，以便它可以集成到现有的荧光或共焦显微镜中，从而显著降低成本，同时最大限度地利用我们现有的基础设施。Quorum的Mizar Tilt是唯一可以添加到标准荧光显微镜上的光片系统，可以将现有系统转换为光片显微镜。通过使用现有的显微镜作为基础，获得薄片显微镜能力的成本显著降低，使Mizar倾斜系统成为最经济且仍具有完整功能的薄片成像解决方案。*联合申请者小组由三个系(工程、科学和健康科学)的工程师、生物学家和化学家组成，正在探索基础和应用研究主题，所有这些主题都受到获得光片显微镜的强烈影响。此外，我们的团队在组织清理和定制细胞培养平台方面拥有特殊的专业知识，使我们能够最大限度地利用这项技术。目前，80多名HQP正在联合申请的实验室中接受培训，其中大多数将能够利用光片显微镜。尽管光片显微镜尚未在工业中广泛使用，但它很可能在几年后成为主流仪器，届时通过收购该仪器而实现的高级培训将使我们的受训人员具有非凡的竞争力，他们的技能将成为生物技术行业雇主梦寐以求的。

As biofabrication and tissue engineering techniques have rapidly advanced in recent years, we are increasingly tasked to visualize complex engineered tissues and 3D biological explants with increasing levels of detail to keep up with the expanding frontier of biological inquiries. However, conventional confocal microscopy is extremely slow in scanning large 3D tissue samples and is unable to track live 3D biological processes in real time. These limitations can be overcome by light-sheet microscopy, which illuminates the tissue sample with a sheet of light to ensure that signal arises only from in-focus regions. Such a mechanism reduces the total sample exposure and increases the scan speed by multiple orders of magnitude, enabling long-term real-time imaging of complex tissues. While our co-applicant group has made significant progress in the development of tissue engineering, organ-on-chip, bioprinting, micro-organism modeling, and drug delivery, we are now severely limited by our ability to characterize and study our tissue samples in 3D. The requested light-sheet microscope is thus essential for probing key characterization questions and enabling novel biological inquiries.******Owing to the rarity of this emerging technology, the use of light-sheet microscopy in tissue engineering and organ-on-a-chip is still in its infancy. To adopt the technique into our collaborative lab space, the light-sheet system should be modular such that it could be integrated into an existing fluorescent or confocal microscope, significantly reducing the cost while maximizing the utility of our existing infrastructure. Quorum's Mizar TILT is the only light-sheet system that can be added to a standard fluorescent microscope to convert an existing system into a light-sheet microscope. By using an existing microscope as a base, the cost of acquiring light sheet microscopy capability is significantly less, making the Mizar TILT system the most economical while still fully functional solution to light-sheet imaging. ******The co-applicant group, composed of a combination engineers, biologists, and chemists across three Faculties (Engineering, Science, and Health Science), is exploring both fundamental and applied research topics, all of which are strongly impacted by the acquisition of a light-sheet microscope. In addition, our team has particular expertise in tissue clearing and customizing cell culture platforms that will enable us to use the technique to its fullest extent. Over 80 HQP are currently being trained among the co-applicant labs, the majority of which would be able to leverage the light sheet microscope. Although the light-sheet microscope is not yet widely used in industry it is likely to become a mainstream instrument in a few years, at which point the advanced training enabled via the acquisition of the instrument will make our trainees exceptionally competitive and their skills highly coveted by employers in the biotechnology sector.*****