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

J Biomed Opt. 2020 Oct;25(10):106504. doi: 10.1117/1.JBO.25.10.106504.

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.

Keywords: axial scan; cylindrical lens; light sheet microscopy; phased array.

MeSH terms

  • Imaging, Three-Dimensional
  • Micro-Electrical-Mechanical Systems*
  • Microscopy*