Open AccessCorrection of depth-induced spherical aberration for deep observation using two-photon excitation fluorescence microscopy with spatial light modulator
Author(s) -
Naoya Matsumoto,
Takashi Inoue,
Akiyuki Matsumoto,
Satoshi Okazaki
Publication year - 2015
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.6.002575
Subject(s) - optics , fluorescence , microscopy , two photon excitation microscopy , materials science , fluorescence microscope , microscope , spatial light modulator , refractive index , fluorescence lifetime imaging microscopy , spherical aberration , light sheet fluorescence microscopy , resolution (logic) , image resolution , physics , artificial intelligence , computer science , lens (geology)
We demonstrate fluorescence imaging with high fluorescence intensity and depth resolution in which depth-induced spherical aberration (SA) caused by refractive-index mismatch between the medium and biological sample is corrected. To reduce the impact of SA, we incorporate a spatial light modulator into a two-photon excitation fluorescence microscope. Consequently, when fluorescent beads in epoxy resin were observed with this method of SA correction, the fluorescence signal of the observed images was ∼27 times higher and extension in the direction of the optical axes was ∼6.5 times shorter at a depth of ∼890 μm. Thus, the proposed method increases the depth observable at high resolution. Further, our results show that the method improved the fluorescence intensity of images of the fluorescent beads and the structure of a biological sample.