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12-15 August 2018
Europe/Berlin timezone

Single- and multi-photon shaped illumination for light-sheet fluorescence microscopy

13 Aug 2018, 10:15


Pfotenhauerstraße 108 01307 Dresden Germany
Short Talk Light sheet fluorescence microscopy Light sheet hardware 1


Dr Jonathan Nylk (University of St Andrews)


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 (e.g. [1]).

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 [2].

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 [3].

Recent progress in shaping optical fields for LSFM will be presented.

[1] T. Vettenburg et al, Nat. Methods 11, 541-544 (2014), doi:10.1038/nmeth.2922
[2] J. Nylk et al, Sci. Adv. 4, eaar4817 (2018), doi:10.1126/sciadv.aar4817
[3] A. Escobet-Montalbán et al, bioRxiv 323790 (2018), doi: 10.1101/323790

Affiliation University of St Andrews
Terms and Conditions Yes

Primary authors

Dr Jonathan Nylk (University of St Andrews) Mr Adrià Escobet-Montalbán (University of St Andrews) Mr Pengfei Liu (University of St Andrews)


Dr Federico Gasparoli (University of St Andrews) Dr Kaley McCluskey (University of St Andrews) Dr Miguel Preciado (University of St Andrews) Dr Michael Mazilu (University of St Andrews) Dr Zhengyi Yang (University of St Aandrews) Prof. Frank Gunn-Moore (University of St Andrews) Ms Sanya Aggarwal (University of St Andrews) Dr Javier Tello (University of St Andrews) Prof. David Ferrier (University of St Andrews) Prof. Kishan Dholakia (University of St Andrews)

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