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Multi-photon attenuation-compensated light-sheet fluorescence microscopy.

ABSTRACT: Attenuation of optical fields owing to scattering and absorption limits the penetration depth for imaging. Whilst aberration correction may be used, this is difficult to implement over a large field-of-view in heterogeneous tissue. Attenuation-compensation allows tailoring of the maximum lobe of a propagation-invariant light field and promises an increase in depth penetration for imaging. Here we show this promising approach may be implemented in multi-photon (two-photon) light-sheet fluorescence microscopy and, furthermore, can be achieved in a facile manner utilizing a graded neutral density filter, circumventing the need for complex beam shaping apparatus. A "gold standard" system utilizing a spatial light modulator for beam shaping is used to benchmark our implementation. The approach will open up enhanced depth penetration in light-sheet imaging to a wide range of end users.

SUBMITTER: Veettikazhy M

PROVIDER: S-EPMC7229186 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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Multi-photon attenuation-compensated light-sheet fluorescence microscopy.

Veettikazhy Madhu M Nylk Jonathan J Gasparoli Federico F Escobet-Montalbán Adrià A Hansen Anders Kragh AK Marti Dominik D Andersen Peter Eskil PE Dholakia Kishan K

Scientific reports 20200515 1

Attenuation of optical fields owing to scattering and absorption limits the penetration depth for imaging. Whilst aberration correction may be used, this is difficult to implement over a large field-of-view in heterogeneous tissue. Attenuation-compensation allows tailoring of the maximum lobe of a propagation-invariant light field and promises an increase in depth penetration for imaging. Here we show this promising approach may be implemented in multi-photon (two-photon) light-sheet fluorescenc ...[more]

PMID: 32415135

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