Propagating-path uniformly scanned light sheet excitation microscopy for isotropic volumetric imaging of large specimens

Journal of Biomedical Optics
Junyu PingPeng Fei

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.

References

Feb 1, 2008·Physical Review Letters·G A SiviloglouD N Christodoulides
Jan 19, 2012·Nature Communications·Florian O Fahrbach, Alexander Rohrbach
Jun 5, 2012·Nature Methods·Uros KrzicLars Hufnagel
Oct 6, 2012·Optics Express·Eugen Baumgart, Ulrich Kubitscheck
Mar 26, 2013·Nature Methods·Misha B AhrensPhilipp J Keller
Apr 8, 2014·Nature Methods·Tom VettenburgKishan Dholakia
Sep 27, 2014·Cell Research·Weijian ZongLiangyi Chen
Nov 18, 2014·Optics Express·Kevin M Dean, Reto Fiolka
Apr 22, 2015·Scientific Reports·Aleksander K ChmielewskiClemens F Kaminski
Apr 30, 2015·Nature Methods·Sébastien WolfRaphaël Candelier
Jun 18, 2015·Biophysical Journal·Kevin M DeanReto Fiolka
Mar 29, 2016·The Journal of Clinical Investigation·Juhyun LeeTzung K Hsiai
Apr 1, 2017·Nature Methods·Rory M Power, Jan Huisken
Mar 18, 2019·BioRxiv : the Preprint Server for Biology·Fabian VoigtF. Helmchen

❮ Previous
Next ❯

Citations

Dec 29, 2020·Journal of the Optical Society of America. A, Optics, Image Science, and Vision·Suhui DengHuilin Zhou
May 18, 2021·Biomedical Optics Express·Bumju KimKi Hean Kim

❮ Previous
Next ❯

Methods Mentioned

BETA
fluorescence microscopy

Software Mentioned

Vignette
PULSE
ORCA
- Flash
ZEMAX

Related Concepts

Related Feeds

Cell Imaging in CNS

Here is the latest research on cell imaging and imaging modalities, including light-sheet microscopy, in the central nervous system.