Multi-photon attenuation-compensated light-sheet fluorescence microscopy
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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.
Veettikazhy , M , Nylk , J , Gasparoli , F , Escobet-Montalbán , A , Hansen , A K , Marti , D , Andersen , P E & Dholakia , K 2020 , ' Multi-photon attenuation-compensated light-sheet fluorescence microscopy ' , Scientific Reports , vol. 10 , 8090 . https://doi.org/10.1038/s41598-020-64891-8
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DescriptionWe thank the UK Engineering and Physical Sciences Research Council for funding (grants EP/P030017/1 and EP/R004854/1), the European Union’s Horizon 2020 Framework Programme (H2020) (675512, BE-OPTICAL), the Danish Council for Independent Research (DFF FTP grant 7017-00021), and the Otto Mønsted Foundation (grant 19-70-0109).
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