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

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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.*****