Abstract

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-photodamage 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 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.

© 2018 Optical Society of America

Light-sheet microscopy: a tutorial

Omar E. Olarte, Jordi Andilla, Emilio J. Gualda, and Pablo Loza-Alvarez
Adv. Opt. Photon. 10(1) 111-179 (2018)

Three-photon fluorescence microscopy with an axially elongated Bessel focus

Cristina Rodríguez, Yajie Liang, Rongwen Lu, and Na Ji
Opt. Lett. 43(8) 1914-1917 (2018)

Homographically generated light sheets for the microscopy of large specimens

Craig T. Russell, Eric J. Rees, and Clemens F. Kaminski
Opt. Lett. 43(4) 663-666 (2018)

Multimodal high-resolution embryonic imaging with light sheet fluorescence microscopy and optical coherence tomography

Behzad Khajavi, Ruijiao Sun, Harshdeep Singh Chawla, Henry H. Le, Manmohan Singh, Alexander W. Schill, Mary E. Dickinson, David Mayerich, and Kirill V. Larin
Opt. Lett. 46(17) 4180-4183 (2021)

Light sheet fluorescence microscopy using high-speed structured and pivoting illumination

Ryosuke Itoh, Joseph Russell Landry, Stephen Sanborn Hamann, and Olav Solgaard
Opt. Lett. 41(21) 5015-5018 (2016)