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Increasing the field-of-view in oblique plane microscopy via optical tiling

2022; Optica Publishing Group; Volume: 13; Issue: 11 Linguagem: Inglês

10.1364/boe.467969

2156-7085

Bingying Chen, Bo-Jui Chang, Felix Zhou, Stephan Daetwyler, Etai Sapoznik, Benjamin A. Nanes, Isabella Terrazas, Gabriel M. Gihana, Lizbeth Perez Castro, Isaac S. Chan, Maralice Conacci‐Sorrell, Kevin M. Dean, Alfred Millett-Sikking, Andrew G. York, Reto Fiolka,

Digital Holography and Microscopy

Fast volumetric imaging of large fluorescent samples with high-resolution is required for many biological applications. Oblique plane microscopy (OPM) provides high spatiotemporal resolution, but the field of view is typically limited by its optical train and the pixel number of the camera. Mechanically scanning the sample or decreasing the overall magnification of the imaging system can partially address this challenge, albeit by reducing the volumetric imaging speed or spatial resolution, respectively. Here, we introduce a novel dual-axis scan unit for OPM that facilitates rapid and high-resolution volumetric imaging throughout a volume of 800 × 500 × 200 microns. This enables us to perform volumetric imaging of cell monolayers, spheroids and zebrafish embryos with subcellular resolution. Furthermore, we combined this microscope with a multi-perspective projection imaging technique that increases the volumetric interrogation rate to more than 10 Hz. This allows us to rapidly probe a large field of view in a dimensionality reduced format, identify features of interest, and volumetrically image these regions with high spatiotemporal resolution.