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Multi-photon attenuation-compensated light-sheet fluorescence microscopy

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 p...

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Detalles Bibliográficos
Autores principales: Veettikazhy, Madhu, Nylk, Jonathan, Gasparoli, Federico, Escobet-Montalbán, Adrià, Hansen, Anders Kragh, Marti, Dominik, Andersen, Peter Eskil, Dholakia, Kishan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7229186/
https://www.ncbi.nlm.nih.gov/pubmed/32415135
http://dx.doi.org/10.1038/s41598-020-64891-8
Descripción
Sumario: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.