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

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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].

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