Scanless confocal light sheet microscopy for high-content and high-throughput imaging of cells and microorganisms

Fluorescent Confocal Microscopy (FCM) has revolutionized biology and medicine by enabling high-resolution, three-dimensional imaging of cells and tissues, advancing our understanding of disease mechanisms and facilitating the development of targeted therapies. However, effective utilization of FCM in many fields is limited by photobleaching/phototoxicity and its relatively slow imaging speed due to point-by-point scanning. Here we report on development of a scanless confocal microscopy technique with light sheet excitation. The core innovation behind this work is utilization of the broadband nature of fluorescence expression to eliminate the need for scanning light across the sample which, in return, significantly lowers the instrumentation complexity and the cost of the system. With such an approach, scanless performance is achieved by taking advantage of a narrow slit (optical encoding in one sample direction) and spectral encoding of the broadband fluorescent emission (spectral encoding in other sample direction). Results from Zemax numerical modeling, experimental characterization tests, and confocal imaging of HeLa and HEK293 cells will be presented.