Browsing by Author "Hu, Yvonne Yuling"

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Temporal focusing-based multiphoton excitation fluorescence images with background noise cancellation via Hilbert-Huang transform
(2019-01-01)
Hu, Yvonne Yuling
;
Luo, Yuan-Rong
;
Lin, Chun-Yu
;
Chang, Chia-Yuan
;
Chen, Shean-Jen
Temporal focusing multiphoton excitation microscopy has wide field-of-view and optical sectioning. By using digital micromirror device, it provides patterned illumination. However, without filling the back aperture of objective lens, the axial confinement is limited to micron-meters, leading the out-of-focus fluorophores excited and image blurred. In this study, Hilbert-Huang transform is proposed to reduce the background noise. The empirical mode decomposition is first applied to disassemble the image into intrinsic mode functions and then reconstruct by Hilbert transform after diminishing background residues. The axial confinement can be enhanced from 2.79 mu m to 0.73 mu m with structure frequency in 1.06 mu m(-1).
Temporal focusing-based widefield multiphoton microscopy with spatially modulated illumination for biotissue imaging
(2018-01-01)
Chang, Chia-Yuan
;
Lin, Cheng-Han
;
Lin, Chun-Yu
;
Sie, Yong-Da
;
Hu, Yvonne Yuling
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. Herein, the TFMPEM has been extended to implement spatially modulated illumination at structured frequency and orientation 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 mm to 1.5 mm for a 50% improvement. By using the TFMPEM with HiLo technique as two structured illuminations at the same spatial frequency but different orientation, reconstructed biotissue images according to the condensed AEC structured illumination are shown obviously superior in contrast and better scattering suppression. Picture: TPEF images of the eosin-stained mouse cerebellar cortex by conventional TFMPEM (left), and the TFMPEM with HiLo technique as 1.09 mm(-1) spatially modulated illumination at 908 (center) and 08 (right) orientations. [GRAPHICS] .