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Probing neural tissue with airy light-sheet microscopy: investigation of imaging performance at depth within turbid media

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Jonathan Nylk, Kaley McCluskey, Sanya Aggarwal, Javier A. Tello, Kishan Dholakia

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Light-sheet microscopy (LSM) has received great interest for fluorescent imaging applications in biomedicine as it facilitates three-dimensional visualisation of large sample volumes with high spatiotemporal resolution whilst minimising irradiation of, and photo-damage to the specimen. Despite these advantages, LSM can only visualize superficial layers of turbid tissues, such as mammalian neural tissue. Propagation-invariant light modes have played a key role in the development of high-resolution LSM techniques as they overcome the natural divergence of a Gaussian beam, enabling uniform and thin light-sheets over large distances. Most notably, Bessel and Airy beam-based light-sheet imaging modalities have been demonstrated. In the single-photon excitation regime and in lightly scattering specimens, Airy-LSM has given competitive performance with advanced Bessel-LSM techniques. Airy and Bessel beams share the property of self-healing, the ability of the beam to regenerate its transverse beam profile after propagation around an obstacle. Bessel-LSM techniques have been shown to increase the penetration-depth of the illumination into turbid specimens but this effect has been understudied in biologically relevant tissues, particularly for Airy beams. It is expected that Airy-LSM will give a similar enhancement over Gaussian-LSM. In this paper, we report on the comparison of Airy-LSM and Gaussian-LSM imaging modalities within cleared and non-cleared mouse brain tissue. In particular, we examine image quality versus tissue depth by quantitative spatial Fourier analysis of neural structures in virally transduced fluorescent tissue sections, showing a three-fold enhancement at 50 μm depth into non-cleared tissue with Airy-LSM. Complimentary analysis is performed by resolution measurements in bead-injected tissue sections.


Original languageEnglish
Title of host publicationThree-Dimensional and Multidimensional Microscopy
Subtitle of host publicationImage Acquisition and Processing XXIV
EditorsThomas G. Brown, Carol J. Cogswell, Tony Wilson
Number of pages7
ISBN (Electronic)9781510605824
ISBN (Print)9781510605817
Publication statusPublished - 17 Feb 2017
EventThree-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIV 2017 - San Francisco, United States
Duration: 30 Jan 20171 Feb 2017
Conference number: 24

Publication series

NameProceedings of SPIE
PublisherSociety of Photo-optical Instrumentation Engineers
ISSN (Print)1605-7422
ISSN (Electronic)2410-9045


ConferenceThree-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIV 2017
CountryUnited States
CitySan Francisco

    Research areas

  • Light-sheet microscopy, LSM, Airy beam, Tissue imaging, Turbid media, Neuroscience

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ID: 262481486