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Propagating-path uniformly scanned light sheet excitation microscopy for isotropic volumetric imaging of large specimens
- Source :
- Journal of Biomedical Optics
- Publication Year :
- 2019
- Publisher :
- Society of Photo-Optical Instrumentation Engineers, 2019.
-
Abstract
- We demonstrate a propagating-path uniformly scanned light sheet excitation (PULSE) microscopy based on the oscillation of voice coil motor that can rapidly drive a thin light sheet along its propagation direction. By synchronizing the rolling shutter of a camera with the motion of laser sheet, we can obtain a uniform plane-illuminated image far beyond the confocal range of Gaussian beam. A stable 1.7-μm optical sectioning under a 3.3 mm×3.3 mm wide field of view (FOV) has been achieved for up to 20 Hz volumetric imaging of large biological specimens. PULSE method transforms the extent of plane illumination from one intrinsically limited by the short confocal range (μm scale) to one defined by the motor oscillation range (mm scale). Compared to the conventional Gaussian light sheet imaging, our method greatly mitigates the compromise of axial resolution and successfully extends the FOV over 100 times. We demonstrate the applications of PULSE method by rapidly imaging cleared mouse spinal cord and live zebrafish larva at isotropic subcellular resolution.
- Subjects :
- Paper
Optics and Photonics
Materials science
Optical sectioning
Light
Biomedical Engineering
Normal Distribution
imaging systems
01 natural sciences
law.invention
optical design
010309 optics
Biomaterials
Mice
Optics
Imaging, Three-Dimensional
law
Oscillometry
0103 physical sciences
Microscopy
Image Processing, Computer-Assisted
Animals
Image resolution
isotropic resolution
Zebrafish
three-dimensional reconstruction
business.industry
Resolution (electron density)
Rolling shutter
Laser
Atomic and Molecular Physics, and Optics
Electronic, Optical and Magnetic Materials
biomedical optics
Microscopy, Fluorescence
Spinal Cord
Light sheet fluorescence microscopy
business
light-sheet microscopy
Algorithms
Gaussian beam
Subjects
Details
- Language :
- English
- ISSN :
- 15602281 and 10833668
- Volume :
- 24
- Issue :
- 8
- Database :
- OpenAIRE
- Journal :
- Journal of Biomedical Optics
- Accession number :
- edsair.doi.dedup.....9ab4d7635c4fd8abb77dcb56d14ead7a