Three-dimensional live multi-label light-sheet imaging with synchronous excitation-multiplexed structured illumination
AffiliationUniv Arizona, Coll Opt Sci
Univ Arizona, Dept Mol & Cell Biol
MetadataShow full item record
PublisherOPTICAL SOC AMER
CitationThree-dimensional live multi-label light-sheet imaging with synchronous excitation-multiplexed structured illumination 2017, 25 (25):31159 Optics Express
Rights© 2017 Optical Society of America under the terms of the OSA Open Access Publishing Agreement
Collection InformationThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at firstname.lastname@example.org.
AbstractMultiplexed imaging is a powerful tool for studying complex interactions inside biological systems. Spectral imaging methods that capture multiple fluorescent markers synchronously without sacrificing the imaging speed or resolution are most suitable for live imaging. We describe spectral-encoded structured illumination (spectral-SIM) light-sheet microscopy, which enables parallel multi-excitation-channel imaging in 3D. Spectral-SIM encodes the excitation wavelength as the phase of the illumination pattern, and allows synchronous image capture over multiple excitation channels at the same speed and spatial resolution as mono-channel structured light-sheet imaging. The technique retains structured light-sheet microscopy's ability in removing out-of-focus and scattered emission background, and generates clear 3D multiplexed images in thick tissue. The capability of this technique was demonstrated by the imaging of live triple-labeled transgenic zebrafish to over 300 mu m deep with 0.5 mu m-by-2 mu m (lateral-by-axial) resolution. (C) 2017 Optical Society of America under the terms of the OSA Open Access Publishing Agreement
NoteOpen access journal.
VersionFinal published version
SponsorsNational Institutes of Health (NIH) [R21EB012646, R01EB015481]