Chow, D.J.X.Schartner, E.P.Corsetti, S.Upadhya, A.Morizet, J.Gunn-Moore, F.J.Dunning, K.R.Dholakia, K.2025-01-062025-01-062024Scientific Reports, 2024; 14(1):20760-1-20760-122045-23222045-2322https://hdl.handle.net/2440/143489Embryo quality assessment by optical imaging is increasing in popularity. Among available optical techniques, light sheet microscopy has emerged as a superior alternative to confocal microscopy due to its geometry, enabling faster image acquisition with reduced photodamage to the sample. However, previous assessments of photodamage induced by imaging may have failed to measure more subtle impacts. In this study, we employed DNA damage as a sensitive indicator of photodamage. We use light sheet microscopy with excitation at a wavelength of 405 nm for imaging embryo autofluorescence and compare its performance to laser scanning confocal microscopy. At an equivalent signal-to-noise ratio for images acquired with both modalities, light sheet microscopy reduced image acquisition time by ten-fold, and did not induce DNA damage when compared to non-imaged embryos. In contrast, imaging with confocal microscopy led to significantly higher levels of DNA damage within embryos and had a higher photobleaching rate. Light sheet imaging is also capable of inducing DNA damage within the embryo but requires multiple cycles of volumetric imaging. Collectively, this study confirms that light sheet microscopy is faster and safer than confocal microscopy for imaging live embryos, indicating its potential as a label-free diagnostic for embryo quality.en© The Author(s) 2024. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. Te images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.Biological techniques; Optics and photonics; PhysicsAnimalsMiceDNA DamageMicroscopy, ConfocalFemaleEmbryo, MammalianOptical ImagingJournal article10.1038/s41598-024-71443-x706960Chow, D.J.X. [0000-0002-2648-4600]Schartner, E.P. [0000-0003-1669-4302]Upadhya, A. [0000-0003-0841-303X]Dunning, K.R. [0000-0002-0462-6479]Dholakia, K. [0000-0001-6534-9009]