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

This work was supported by awards to KD from the UK Engineering and Physical Sciences Research Council under grant EP/J01771X/1, the ’BRAINS’ 600th anniversary appeal, and Dr. E. Killick. We would also like to thank The Northwood Trust and The RS Macdonald Charitable Trust for funding support (awarded to JAT). KD acknowledges support of a Royal Society Leverhulme Trust Senior Fellowship.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.Publisher PDFPeer reviewe