Multi-photon attenuation-compensated light-sheet fluorescence microscopy

Madhu Veettikazhy (Lead / Corresponding author), Jonathan Nylk, Federico Gasparoli, Adrià Escobet-Montalbán, Anders Kragh Hansen, Dominik Marti, Peter Eskil Andersen, Kishan Dholakia

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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.

Original languageEnglish
Article number8090
Pages (from-to)1-10
Number of pages10
JournalScientific Reports
Volume10
DOIs
Publication statusPublished - 15 May 2020

Keywords

  • Cellular imaging
  • Light-sheet microscopy
  • Microscopy
  • Multiphoton microscopy
  • Optics and photonics

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    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, 10, 1-10. [8090]. https://doi.org/10.1038/s41598-020-64891-8