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

用于工程组织和微生理系统 3D 生物成像的光片显微镜

• 批准号: RTI-2020-00463
• 负责人: Zhang, Boyang
• 金额: $ 10.92万
• 依托单位: McMaster University
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=693690
• 关键词: Light; sheet; microscopy; 3D; bioimaging

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.*****

近年来，随着生物制造和组织工程技术的迅速发展，我们越来越多地承担着可视化复杂工程组织和3D生物外植体的任务，其细节水平不断提高，以跟上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组织样本时非常慢，并且不能真实的跟踪活的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，以确保信号仅来自聚焦区域。这种机制减少了总的样品曝光，并将扫描速度提高了多个数量级，从而能够对复杂组织进行长期实时成像。虽然我们的共同申请人小组在组织工程、芯片上器官、生物打印、微生物建模和药物递送方面取得了重大进展，但我们现在在3D中表征和研究组织样本的能力受到严重限制。因此，所需的光片显微镜对于探索关键表征问题和实现新的生物学研究至关重要。由于这种新兴技术的稀有性，光片显微镜在组织工程和器官芯片中的应用仍处于起步阶段。 为了将该技术应用于我们的协作实验室空间，光片系统应该是模块化的，这样它就可以集成到现有的荧光或共聚焦显微镜中，从而大大降低成本，同时最大限度地利用我们现有的基础设施。Quorum的Mizar TILT是唯一一个可以添加到标准荧光显微镜中的光片系统，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，获得光片显微镜功能的成本显着降低，使Mizar TILT系统成为最经济，同时仍然功能齐全的光片成像解决方案。** 共同申请人小组由三个学院（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课题，所有这些都受到收购光片显微镜的强烈影响。此外，我们的团队在组织清除和定制细胞培养平台方面具有特殊的专业知识，这将使我们能够最大限度地使用该技术。 超过80名HQP目前正在共同申请人实验室中接受培训，其中大多数将能够利用光片显微镜。虽然光片显微镜尚未在工业中广泛使用，但它很可能在几年内成为主流仪器，届时通过购买该仪器实现的高级培训将使我们的学员极具竞争力，他们的技能受到生物技术领域雇主的高度垂涎。