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

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

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

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, Tony Wilson, Carol J. Cogswell
Place of PublicationBellingham, WA
PublisherSPIE-International Society for Optical Engineering
Number of pages7
Volume10070
ISBN (Electronic)9781510605824
ISBN (Print)9781510605817
DOIs
Publication statusPublished - 2017
EventThree-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIV 2017 - San Francisco, United States
Duration: 30 Jan 20171 Feb 2017

Publication series

NameProceedings of SPIE
ISSN (Print)1605-7422
ISSN (Electronic)2410-9045

Conference

ConferenceThree-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIV 2017
CountryUnited States
CitySan Francisco
Period30/01/171/02/17

Keywords

  • Airy beam
  • Light-sheet microscopy
  • LSM
  • neuroscience
  • Tissue imaging
  • turbid media

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  • Cite this

    Nylk, J., McCluskey, K., Aggarwal, S., Tello, J. A., & Dholakia, K. (2017). Probing neural tissue with airy light-sheet microscopy: Investigation of imaging performance at depth within turbid media. In T. G. Brown, T. Wilson, & C. J. Cogswell (Eds.), Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIV (Vol. 10070). [100700B] (Proceedings of SPIE). SPIE-International Society for Optical Engineering. https://doi.org/10.1117/12.2251921