PUBLICATION
Propagating-path uniformly scanned light sheet excitation microscopy for isotropic volumetric imaging of large specimens
- Authors
- Ping, J., Zhao, F., Nie, J., Yu, T., Zhu, D., Liu, M., Fei, P.
- ID
- ZDB-PUB-190807-18
- Date
- 2019
- Source
- Journal of Biomedical Optics 24: 1-5 (Journal)
- Registered Authors
- Liu, Mugen, Zhao, Feng
- Keywords
- biomedical optics, imaging systems, isotropic resolution, light-sheet microscopy, optical design, three-dimensional reconstruction
- MeSH Terms
-
- Algorithms
- Animals
- Image Processing, Computer-Assisted/methods*
- Imaging, Three-Dimensional
- Light
- Mice
- Microscopy, Fluorescence/methods*
- Normal Distribution
- Optics and Photonics
- Oscillometry
- Spinal Cord/diagnostic imaging*
- Zebrafish
- PubMed
- 31385482 Full text @ J. Biomed. Opt.
Citation
Ping, J., Zhao, F., Nie, J., Yu, T., Zhu, D., Liu, M., Fei, P. (2019) Propagating-path uniformly scanned light sheet excitation microscopy for isotropic volumetric imaging of large specimens. Journal of Biomedical Optics. 24:1-5.
Abstract
<p>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.</p>.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping