Enhancement of image quality and imaging depth with Airy light-sheet microscopy in cleared and non-cleared neural tissue

Abstract

We have investigated the effect of Airy illumination on the image quality and depth penetration of digitally scanned light-sheet microscopy in turbid neural tissue. We used Fourier analysis of images acquired using Gaussian and Airy light-sheets to assess their respective image quality versus penetration into the tissue. We observed a three-fold average improvement in image quality at 50 μm depth with the Airy light-sheet. We also used optical clearing to tune the scattering properties of the tissue and found that the improvement when using an Airy light-sheet is greater in the presence of stronger sample-induced aberrations. Finally, we used homogeneous resolution probes in these tissues to quantify absolute depth penetration in cleared samples with each beam type. The Airy light-sheet method extended depth penetration by 30% compared to a Gaussian light-sheet.

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

“…The FWHM obtained with the Airy was measured to be ∼3.6 µm at the surface and remained quite confined at depth reaching the value of 4.3 µm at 400 µm from the surface. These results agree with what was previously observed by Nylk et al [19]. The Airy light sheet penetrates deeper into the sample without aberration correction.…”

“…Compared to the cleared sample, the depth range in turbid tissue is severely limited by the high scattering. While the advantage of propagation invariance is less evident, the Airy beam has an additional advantage in depth penetration through scattering tissue due to the angular diversity and self healing property of the beam shape [19]. We see this manifest with sharper tissue features and reduced aberrations in depth.…”

“…In this paper, we breach this limit to resolution with the use of deep learning to recover high-resolution images from sub-sampled information. We couple this with the particular use of a propagation-invariant Airy light field for illumination, which allows for a wider field of view captured on the camera than when using a Gaussian beam; furthermore, it has inherent advantages in achieving increased depth penetration through aberrating tissues [19]. In this way, we realise imaging across length scales with LSM over extended fields of view and depth ranges.…”

Light-sheet microscopy: a tutorial

“…Also, just like optical lattices, ALSs excel at confining the excitation to the dimensions perpendicular to the detection path and reducing peak intensities minimizing phototoxicity. Thus, ALSs have been successfully applied to image medium-sized samples, from small cell spheroids to sections of small vertebrates [65] and mice neural tissue [161].…”

Light-Sheet Fluorescence Microscopy with Scanning Non-diffracting Beams

“…The first two approaches although lead to generations of uniform light-sheet in a wider field-of-view (FOV) and mitigates the ghost image, inevitably result in longer exposure of the sample to exciting illumination and therefore may induce photodamage, and also have rather complex experimental arrangements. Using non-diffracting beams such as Bessel beam [13][14][15], modified Bessel beam [16], and Airy beams [17][18][19] have also been proposed for this purpose. Although, the self-healing property of the nondiffracting beams is expected to enhance the uniformity of the illumination light sheet, their mutli-lobe intensity structure leads to simultaneous illumination of various depths, and reduces the image contrast.…”