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Single- and multi-photon shaped illumination for light-sheet fluorescence microscopy (Conference Presentation)
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Paper Abstract

The use of exotic optical modes is becoming increasingly widespread in microscopy. Particularly, propagation-invariant beams, such as Airy and Bessel beams and optical lattices, have been particularly useful in light-sheet fluorescence microscopy (LSFM) as they enable high-resolution imaging over a large field-of-view (FOV), possess a resistance to the deleterious effects of specimen induced light scattering, and can potentially reduce photo-toxicity. Although these propagation-invariant beams can resist the effects of light scattering to some degree, and there has been some interest in adaptive-optical methods to correct for beam aberrations when they cannot, scattering and absorption of the illuminating light-sheet limit the penetration of LSFM into tissues and results in non-uniform intensity across the FOV. A new degree of control over the intensity evolution of propagation-invariant beams can overcome beam losses across the FOV, restoring uniform illumination intensity and therefore image quality. This concept is compatible with all types of propagation-invariant beams and is characterised in the context of light-sheet image quality. Another property to control is the wavelength of light used. Optical transmission through tissue is greatly improved at longer wavelengths into the near-infrared due to reduced Rayleigh scattering and two-photon excitation has proved beneficial for imaging at greater depth in LSFM. Three-photon excitation has already been demonstrated as a powerful tool to increase tissue penetration in deep brain confocal microscopy, and when combined with beam shaping can also be a powerful illumination strategy for LSFM. Recent progress in shaping optical fields for LSFM will be presented.

Paper Details

Date Published: 4 March 2019
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Proc. SPIE 10883, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXVI, 1088319 (4 March 2019); doi: 10.1117/12.2509353
Show Author Affiliations
Jonathan Nylk, Univ. of St. Andrews (United Kingdom)
Adrià Escobet-Montalbán, Univ. of St. Andrews (United Kingdom)
Pengfei Liu, Univ. of St. Andrews (United Kingdom)
Federico M. Gasparoli, Univ. of St. Andrews (United Kingdom)
Kaley McCluskey, Univ. of St Andrews (United Kingdom)
Miguel A. Preciado, Univ. of St. Andrews (United Kingdom)
Michael Mazilu, Univ. of St. Andrews (United Kingdom)
Zhengyi Yang, Univ. of St. Andrews (United Kingdom)
Frank J. Gunn-Moore, Univ. of St. Andrews (United Kingdom)
Sanya Aggarwal, Univ. of St. Andrews (United Kingdom)
Javier A. Tello, Univ. of St. Andrews (United Kingdom)
David E. K. Ferrier, Univ. of St. Andrews (United Kingdom)
Kishan Dholakia, Univ. of St. Andrews (United Kingdom)


Published in SPIE Proceedings Vol. 10883:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXVI
Thomas G. Brown; Tony Wilson, Editor(s)

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