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Proceedings Article

Numerical spherical aberration correction method using spatial light modulator under deep-part fluorescence observation

[+] Author Affiliations
Yu Takiguchi, Hisayoshi Takamoto, Takashi Inoue, Naoya Matsumoto

Hamamatsu Photonics K.K. (Japan)

Masamitsu Kanada, Susumu Terakawa

Hamamatsu Univ. School of Medicine (Japan)

Proc. SPIE 8949, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXI, 89490C (March 12, 2014); doi:10.1117/12.2038394
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From Conference Volume 8949

  • Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXI
  • Thomas G. Brown; Carol J. Cogswell; Tony Wilson
  • San Francisco, California, United States | February 01, 2014

abstract

We have developed a confocal fluorescence laser scanning microscopy (CFLSM) incorporating a liquid crystal on silicon spatial light modulator (LCOS-SLM). To achieve high-resolution and high-contrast imaging for deeper part of the tissue with CFLSM, high numerical aperture objective lenses are required to tightly focus excitation light to meet Rayleigh limit(criterion) for the specimens. However, mismatch of refractive index at the boundary of interfacing materials, such as atmosphere, glass cover, and biological tissues, causes spherical aberration. Recently, we proposed a numerical method for correcting spherical aberration. In this method a pre-distorted wavefront pattern for aberration correction is calculated by ray tracing from a hypothetical focal point inside a specimen to the pupil plane. The resulting microscope can correct such spherical aberration. We observed 6.0μm fluorescent micro-beads dispersed three-dimensionally in agarose gel to confirm effectiveness of aberration correction. We reconstructed a three-dimensional image by taking 20 images by changing the depth with 1 μm interval and stacking them. It was apparent that the longitudinal/depth resolution was improved and that the intensity of fluorescence image was increased with aberration correction. While this method is applicable to other laser scanning microscopes, it has potential to enhance the signals for various super-resolution microscopic techniques, such as stimulated- emission-depletion (STED) fluorescence microscopy. © (2014) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Citation

Yu Takiguchi ; Hisayoshi Takamoto ; Masamitsu Kanada ; Takashi Inoue ; Naoya Matsumoto, et al.
" Numerical spherical aberration correction method using spatial light modulator under deep-part fluorescence observation ", Proc. SPIE 8949, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXI, 89490C (March 12, 2014); doi:10.1117/12.2038394; http://dx.doi.org/10.1117/12.2038394


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