Widefield light sheet microscopy using an Airy beam combined with deep-learning super-resolution

Stella Corsetti,Federico Gasparoli,C Simon Herrington,Shuzo Sakata,Kishan Dholakia,Philip Wijesinghe,Khushi Vyas,Persephone B Poulton,Jonathan Nylk

doi:10.1364/osac.391644

Abstract

Imaging across length scales and in depth has been an important pursuit of widefield optical imaging. This promises to reveal fine cellular detail within a widefield snapshot of a tissue sample. Current advances often sacrifice resolution through selective sub-sampling to provide a wide field of view in a reasonable time scale. We demonstrate a new avenue for recovering high-resolution images from sub-sampled data in light sheet microscopy using deep-learning super-resolution. We combine this with the use of a widefield Airy beam to achieve high-resolution imaging over extended fields of view and depths. We characterise our method on fluorescent beads as test targets. We then demonstrate improvements in imaging amyloid plaques in a cleared brain from a mouse model of Alzheimer’s disease, and in excised healthy and cancerous colon and breast tissues. This development can be widely applied in all forms of light sheet microscopy to provide a two-fold increase in the dynamic range of the imaged length scale. It has the potential to provide further insight into neuroscience, developmental biology, and histopathology.

Highlights

Widefield optical imaging at depth has opened up new vistas for neuroscience and developmental biology [1,2,3], and is expanding its remit into histopathology [4]
The illumination is split into two parallel optical paths, with one path modulated by a one-dimensional cubic polynomial phase mask (PM)
The model in this paper is based on the pre-trained set that maps low resolution (10x, 0.4 numerical aperture (NA)) images to higher-resolution co-registered (20x, 0.75NA) images in widefield epifluorescence microscopy. We found this approach to be well-suited for the light sheet microscopy (LSM) system, for both the Airy and Gaussian illuminations, as we detail in the Results and Discussion sections

Summary

Introduction

Widefield optical imaging at depth has opened up new vistas for neuroscience and developmental biology [1,2,3], and is expanding its remit into histopathology [4]. Recent advances in high-resolution imaging with structured illumination microscopy (SIM) [8,9] or at ultraviolet excitation wavelengths (MUSE) [10] have revealed novel information on the sub-cellular scale; they have often been at the sacrifice of the size of the area probed, expense or imaging speed. The geometry of LSM, with orthogonal illumination and detection arms, has shown a significant reduction in photodamage accompanied with rapid widefield volumetric image acquisition. This has been married with new photonics innovations, such as the use of propagation-invariant light fields, and multi-objective schemes, offering a multiview sample perspective and a high isotropic resolution [11,12].

Methods

Results

Discussion

Conclusion