Open AccessHard-wired lattice light-sheet microscopy for imaging of expanded samples
Author(s) -
Anne Stockhausen,
Jana Bürgers,
Juan Eduardo Rodriguez-Gatica,
Jens F. Schweihoff,
Rudolf Merkel,
Jens Markus Prigge,
Martin K. Schwarz,
Ulrich Kubitscheck
Publication year - 2020
Publication title -
optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.394
H-Index - 271
ISSN - 1094-4087
DOI - 10.1364/oe.393728
Subject(s) - light sheet fluorescence microscopy , microscopy , optics , materials science , microscope , optical microscope , polarized light microscopy , bright field microscopy , depth of field , light field , scanning confocal electron microscopy , physics , scanning electron microscope
Light-sheet fluorescence microscopy (LSFM) helps investigate small structures in developing cells and tissue for three-dimensional localization microscopy and large-field brain imaging in neuroscience. Lattice light-sheet microscopy is a recent development with great potential to improve axial resolution and usable field sizes, thus improving imaging speed. In contrast to the commonly employed Gaussian beams for light-sheet generation in conventional LSFM, in lattice light-sheet microscopy an array of low diverging Bessel beams with a suppressed side lobe structure is used. We developed a facile elementary lattice light-sheet microscope using a micro-fabricated fixed ring mask for lattice light-sheet generation. In our setup, optical hardware elements enable a stable and simple illumination path without the need for spatial light modulators. This setup, in combination with long-working distance objectives and the possibility for simultaneous dual-color imaging, provides optimal conditions for imaging extended optically cleared tissue samples. We here present experimental data of fluorescently stained neurons and neurites from mouse hippocampus following tissue expansion and demonstrate the high homogeneous resolution throughout the entire imaged volume. Utilizing our purpose-built lattice light-sheet microscope, we reached a homogeneous excitation and an axial resolution of 1.2 µm over a field of view of (333 µm) 2 .