Open AccessCorrection of spherical aberration in multi-focal multiphoton microscopy with spatial light modulator
Author(s) -
Naoya Matsumoto,
Alu Konno,
Yasushi Ohbayashi,
Takashi Inoue,
Akiyuki Matsumoto,
Kenji Uchimura,
Kenji Kadomatsu,
Satoshi Okazaki
Publication year - 2017
Publication title -
optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.394
H-Index - 271
ISSN - 1094-4087
DOI - 10.1364/oe.25.007055
Subject(s) - optics , spherical aberration , spatial light modulator , microscopy , materials science , wavefront , focal length , refractive index , fluorescence , fluorescence microscope , distortion (music) , microscope , physics , optoelectronics , lens (geology) , amplifier , cmos
We demonstrate that high-quality images of the deep regions of a thick sample can be obtained from its surface by multi-focal multiphoton microscopy (MMM). The MMM system incorporates a spatial light modulator to separate the excitation beam into a multi-focal excitation beam and modulate the pre-distortion wavefront to correct spherical aberration (SA) caused by a refractive index mismatch between the immersion medium and the biological sample. When fluorescent beads in transparent epoxy resin were observed using four SA-corrected focal beams, the fluorescence signal of the obtained images was ~52 times higher than that obtained without SA correction until a depth of ~1100 μm, similar to the result for single-focal multiphoton microscopy (SMM). The MMM scanning time was four times less than that for SMM, and MMM showed an improved fluorescence intensity and depth resolution for an image of blood vessels in the brain of a mouse stained with a fluorescent dye.