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Lissajous scanning structured illumination microscopy
- Source :
- Biomed Opt Express
- Publication Year :
- 2020
- Publisher :
- Optica Publishing Group, 2020.
-
Abstract
- High-resolution fluorescent microscopic imaging techniques are in high demand to observe detailed structures or dynamic mechanisms of biological samples. Structured illumination microscopy (SIM) has grabbed much attention in super-resolution imaging due to simple configuration, high compatibility with common fluorescent molecules, and fast image acquisition. Here, we report Lissajous scanning SIM (LS-SIM) by using a high fill-factor Lissajous scanning micromirror and laser beam modulation. The LS-SIM was realized by a Lissajous scanned structured illumination module, relay optics, and a conventional fluorescent microscope. The micromirror comprises an inner mirror and an outer frame, which are scanned at pseudo-resonance with electrostatic actuation. The biaxial scanning frequencies are selected by the frequency selection rule for high fill-factor (> 80%) Lissajous scanning. Structured illumination (SI) was then realized by modulating the intensity of a laser beam at the least common multiple (LCM) of the scanning frequencies. A compact Lissajous scanned SI module containing a fiber-optic collimator and Lissajous micromirror has been fully packaged and coupled with relay optics and a fiber-based diode pumped solid state (DPSS) laser including acousto-optic-modulator (AOM). Various structured images were obtained by shifting the phase and orientation of the illumination patterns and finally mounted with a conventional fluorescent microscope. The LS-SIM has experimentally demonstrated high-resolution fluorescent microscopic imaging of reference targets and human lung cancer cell PC-9 cells. The LS-SIM exhibits the observable region in spatial frequency space over 2x, the line-edge sharpness over 1.5x, and the peak-to-valley (P-V) ratio over 2x, compared to widefield fluorescent microscopy. This method can provide a new route for advanced high-resolution fluorescent microscopic imaging.
- Subjects :
- 0303 health sciences
Materials science
business.industry
Structured illumination microscopy
Collimator
Laser
01 natural sciences
Fluorescence
Article
Atomic and Molecular Physics, and Optics
law.invention
010309 optics
Lissajous curve
03 medical and health sciences
Optics
law
0103 physical sciences
Fluorescence microscope
Spatial frequency
business
030304 developmental biology
Biotechnology
Diode
Subjects
Details
- ISSN :
- 21567085
- Volume :
- 11
- Database :
- OpenAIRE
- Journal :
- Biomedical Optics Express
- Accession number :
- edsair.doi.dedup.....126d54e0aa08bdb330e264ba53ce795f