Multidirectional digital scanned light-sheet microscopy enables uniform fluorescence excitation and contrast-enhanced imaging

Scientific Reports
Adam K GlaserJonathan T C Liu

Abstract

Light-sheet fluorescence microscopy (LSFM) has emerged as a powerful method for rapid and optically efficient 3D microscopy. Initial LSFM designs utilized a static sheet of light, termed selective plane illumination microscopy (SPIM), which exhibited shadowing artifacts and deteriorated contrast due to light scattering. These issues have been addressed, in part, by multidirectional selective plane illumination microscopy (mSPIM), in which rotation of the light sheet is used to mitigate shadowing artifacts, and digital scanned light-sheet microscopy (DSLM), in which confocal line detection is used to reject scattered light. Here we present a simple and passive multidirectional digital scanned light-sheet microscopy (mDSLM) architecture that combines the benefits of mSPIM and DSLM. By utilizing an elliptical Gaussian beam with increased angular diversity in the imaging plane, mDSLM provides mitigation of shadowing artifacts and contrast-enhanced imaging of fluorescently labeled samples.

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Citations

Jul 6, 2019·Nature Communications·Adam K GlaserJonathan T C Liu
Feb 23, 2020·Frontiers in Immunology·Samantha J MontagueElizabeth E Gardiner
Jul 9, 2020·Biomedical Optics Express·Pietro RicciFrancesco Saverio Pavone
Feb 17, 2021·Methods and Applications in Fluorescence·Sergiy V Avilov
May 22, 2021·Nature Communications·Sota TakanezawaTakeshi Imamura
Jun 15, 2021·Progress in Biophysics and Molecular Biology·Gregory B SandsBruce H Smaill
Jul 23, 2021·Progress in Biophysics and Molecular Biology·Pablo Delgado-RodriguezArrate Muñoz-Barrutia
Jul 19, 2021·Progress in Biophysics and Molecular Biology·Pietro RicciFrancesco Saverio Pavone

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Methods Mentioned

BETA
fluorescence microscopy
light-sheet
Rayleigh scattering

Software Mentioned

MATLAB
Windows
mDSLM
SPIM
mSPIM
DSLM
LABVIEW
ImageJ

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