High content live imaging on an inverted light-sheet microscope
Au, Aaron 1 ; Yip, Christopher 1, 2, 3
1. Department of Applied Chemistry and Chemical Engineering; 2. Institute of Biomaterials & Biomedical Engineering; 3. Department of Biochemistry
Light sheet microscopy has gained popularity in recent years due to its higher throughput imaging speed at reduced sample phototoxicity compared to conventional confocal microscopy. However in practice, the arduous process of sample mounting often hinders the high-throughput imaging pipeline to be efficacious. In addition, a typical mounting method is restricted to small selections of biological system due to a large variation between sample shapes, mounting medium required, and the geometry of the light sheet platform.
Inspired by the concept from Strnad et.al , we developed a light sheet microscope that allows an adaptable open top sample chamber to be fitted on a confocal microscope base. This allows us to take advantage of the existing confocal laser lines and galvo scanning mirrors to create the scanning light sheet. The additional optics can easily be removed to and the microscope can be converted back to a confocal microscope.
The inverted light sheet configuration provides generous amount of space for large sample holders that can be adopted for a wide range of biological sample sizes and geometries. Combined with our in-house rapid prototyping capabilities, the mounting space can be iteratively redesigned and customized for a plethora of biological system of interest. This has allowed us to prototype sample mounts to perform high-throughput imaging. We demonstrate the versatility of our in house light sheet system with various biological samples in their optimized sample mounts.