High-speed axially swept light sheet microscopy using a linear MEMS phased array for isotropic resolution

Abstract

SIGNIFICANCE: Axially swept light sheet microscopy is used for deconvolution-free, high-resolution 3D imaging, but usually the axial scan mechanism reduces the top imaging speed. Phased arrays (PAs) for axial scanning enable both high resolution and high speed. AIM: A high-speed PA with an update rate faster than the camera row read time is used to track the rolling shutter at camera-limited rates. APPROACH: The point spread function is evaluated to ensure sub-micron isotropic resolution, and the technique is demonstrated on a live Drosophila embryo. RESULTS: Isotropic resolution is shown down to 720  ±  55  nm in all three spatial dimensions. With an update rate of 2.85  μs, the PA tracks the camera sensor rolling shutter at camera-limited rates. Features in the Drosophila embryo are resolved clearly compared with the equivalent static light sheet case. The random-access nature of the PA enables a camera sensor readout in the same direction for each frame to maintain even temporal sampling in image sequences with no speed loss. CONCLUSIONS: Use of PAs is compatible with axially swept light sheet microscopy and offers significant improvements in speed.