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Single-Cell Biophysics: Measurement, Modulation, and Modeling
Saturday Speaker Abstracts
10
Lattice Light Sheet Microscopy on Single-Cell Imaging
Bi-Chang Chen
.
Academia Sinica, Taipei, Taiwan.
Optical imaging techniques are constantly evolving with the desire to innovate an imaging tool
that is capable of seeing sub-cellular processes in a biological system, especailly in three
dimensions. We crafted ultra-thin light sheets derived from two-dimensional optical lattices that
allowed us to image three-dimensional (3D) dynamics for hundreds of volumes, often at sub-
second intervals, at the diffraction limit and beyond. We applied this tool to systems spanning
four orders of magnitude in space and time, including the diffusion of single transcription factor
molecules in a spheroid of stem cells, the 3D dynamic instability of microtubules during mitosis,
the formation of the immunological synapse, neutrophil motility in a 3D matrix, and
embryogenesis in Caenorhabditis elegans and Drosophila melanogaster.
Deep-Biotissue Imaging by Temporal Focusing Widefield Multiphoton Microscopy
Shean-Jen Chen
1
, Chia-Yuan Chang
2,1
, Yong-Da Sie
3
.
1
National Chiao Tung University, Tainan, Taiwan,
2
National Cheng Kung University, Tainan,
Taiwan,
3
National Cheng Kung University, Tainan, Taiwan.
A developed temporal focusing-based multiphoton excitation microscope (TFMPEM) has a
digital micromirror device (DMD) which is adopted not only as a blazed grating for light spatial
dispersion but also for patterned illumination simultaneously. The TFMPEM has been extended
to implement spatially modulated and digital holographic illumination to increase the beam
coverage at the back-focal aperture of the objective lens. The axial excitation confinement (AEC)
of TFMPEM can be condensed from 3.0 μm to 1.5 μm. By using the TFMPEM with HiLo
technique, reconstructed deep-biotissue images according to the condensed AEC structured
illumination are shown obviously superior in contrast and better scattering suppression.