Open AccessTemporal focusing microscopy using three-photon excitation fluorescence with a 92-fs Yb-fiber chirped pulse amplifier
Author(s) -
Keisuke Toda,
Keisuke Isobe,
Kamiki,
Hiroyuki Kawano,
Atsushi Miyawaki,
Katsumi Midorikawa
Publication year - 2017
Publication title -
biomedical optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.362
H-Index - 86
ISSN - 2156-7085
DOI - 10.1364/boe.8.002796
Subject(s) - microscopy , two photon excitation microscopy , materials science , optics , fluorescence microscope , optical sectioning , light sheet fluorescence microscopy , optical microscope , excitation , ultrashort pulse , fluorescence lifetime imaging microscopy , microscope , fluorescence , point spread function , femtosecond , optoelectronics , scanning confocal electron microscopy , laser , physics , scanning electron microscope , quantum mechanics
Temporal focusing (TF) microscopy is a wide-field two-photon excitation fluorescence (2PEF) microscopy technique, the optical sectioning capability of which is lower than that of point-scanning 2PEF microscopy. Here we demonstrate TF microscopy using three-photon excitation fluorescence (3PEF), which enhances the optical sectioning capability. As an excitation light source for the 3PEF, we developed an Yb-fiber chirped pulse amplifier, which produces 92-fs 9.0-μJ 1060-nm pulses at a repetition rate of 200 kHz. The optical sectioning capability was improved by a factor of 1.3 compared with that of 2PEF-TF microscopy. We also demonstrate dual-color imaging with both 2PEF and 3PEF.