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

Jonathan Nylk; Adrià Escobet-Montalbán; Pengfei Liu; Federico M. Gasparoli; Kaley McCluskey; Miguel A. Preciado; Michael Mazilu; Zhengyi Yang; Frank J. Gunn-Moore; Sanya Aggarwal; Javier A. Tello; David E. K. Ferrier; Kishan Dholakia

doi:10.1117/12.2509353

4 March 2019 Single- and multi-photon shaped illumination for light-sheet fluorescence microscopy (Conference Presentation)

Jonathan Nylk, Adrià Escobet-Montalbán, Pengfei Liu, Federico M. Gasparoli, Kaley McCluskey, Miguel A. Preciado, Michael Mazilu, Zhengyi Yang, Frank J. Gunn-Moore, Sanya Aggarwal, Javier A. Tello, David E. K. Ferrier, Kishan Dholakia

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Proceedings Volume 10883, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXVI; 1088319 (2019) https://doi.org/10.1117/12.2509353
Event: SPIE BiOS, 2019, San Francisco, California, United States

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.

Conference Presentation

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Jonathan Nylk, Adrià Escobet-Montalbán, Pengfei Liu, Federico M. Gasparoli, Kaley McCluskey, Miguel A. Preciado, Michael Mazilu, Zhengyi Yang, Frank J. Gunn-Moore, Sanya Aggarwal, Javier A. Tello, David E. K. Ferrier, and Kishan Dholakia "Single- and multi-photon shaped illumination for light-sheet fluorescence microscopy (Conference Presentation)", Proc. SPIE 10883, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXVI, 1088319 (4 March 2019); https://doi.org/10.1117/12.2509353

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KEYWORDS

Microscopy

Multiphoton microscopy

Luminescence

Multiphoton fluorescence microscopy

Light scattering

Tissues

Beam propagation method

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