Three-photon light-sheet fluorescence microscopy

We present the first demonstration of three-photon excitation light-sheet fluorescence microscopy. Light-sheet fluorescence microscopy in single- and two-photon modes has emerged as a powerful wide-field, low photo-damage technique for fast volumetric imaging of biological samples. We extend this imaging modality to the three-photon regime enhancing its penetration depth. Our present study uses a standard conventional femtosecond pulsed laser at 1000 nm wavelength for the imaging of 450 μm diameter cellular spheroids. In addition, we show, experimentally and through numerical simulations, the potential advantages in three-photon light-sheet microscopy of using propagation-invariant Bessel beams in preference to Gaussian beams.

Citation

Escobet Montalban , A , Gasparoli , F M , Nylk , J , Liu , P , Yang , Z & Dholakia , K 2018 , ' Three-photon light-sheet fluorescence microscopy ' , Optics Letters , vol. 43 , no. 21 , pp. 5484-5487 . https://doi.org/10.1364/OL.43.005484

Publication

Optics Letters

Status

Peer reviewed

DOI

https://doi.org/10.1364/OL.43.005484

ISSN

0146-9592

Type

Journal article

Rights

Copyright © 2018 Optical Society of America. This work has been made available online in accordance with the publisher’s policies. This is the author created, accepted version manuscript following peer review and may differ slightly from the final published version. The final published version of this work is available at: https://doi.org/10.1364/OL.43.005484

Description

This work has received funding from the European Union’s Horizon 2020 Programme through the project Advanced BiomEdical OPTICAL Imaging and Data Analysis (BEOPTICAL) under grant agreement no. 675512 and the UK Engineering and Physical Sciences Research Council (grants EP/P030017/1 and EP/R004854/1).