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.
Citation
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
Publication
Scientific Reports
Status
Peer reviewed
ISSN
2045-2322Type
Journal article
Description
We 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).Collections
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