In vivo three- and four-photon fluorescence microscopy using a 1.8 µm femtosecond fiber laser system.
Biomed Opt Express
; 14(1): 326-334, 2023 Jan 01.
Article
en En
| MEDLINE
| ID: mdl-36698657
ABSTRACT
Multiphoton microscopy has enabled us to image cellular dynamics in vivo. However, the excitation wavelength for imaging with commercially available lasers is mostly limited between 0.65-1.04 µm. Here we develop a femtosecond fiber laser system that produces â¼150 fs pulses at 1.8 µm. Our system starts from an erbium-doped silica fiber laser, and its wavelength is converted to 1.8 µm using a Raman shift fiber. The 1.8 µm pulses are amplified with a two-stage TmZBLAN fiber amplifier. The final pulse energy is â¼1 µJ, sufficient for in vivo imaging. We successfully observe TurboFP635-expressing cortical neurons at a depth of 0.7 mm from the brain surface by three-photon excitation and Clover-expressing astrocytes at a depth of 0.15 mm by four-photon excitation.
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MEDLINE
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En
Revista:
Biomed Opt Express
Año:
2023
Tipo del documento:
Article
País de afiliación:
Japón