Abstract
Light Sheet Fluorescence Microscopy (LSFM) enables multi-dimensional and multi-scale imaging via illuminating specimens with a separate thin sheet of laser. It allows rapid plane illumination for reduced photo-damage and superior axial resolution and contrast. We hereby demonstrate cardiac LSFM (c-LSFM) imaging to assess the functional architecture of zebrafish embryos with a retrospective cardiac synchronization algorithm for four-dimensional reconstruction (3-D space + time). By combining our approach with tissue clearing techniques, we reveal the entire cardiac structures and hypertrabeculation of adult zebrafish hearts in response to doxorubicin treatment. By integrating the resolution enhancement technique with c-LSFM to increase the resolving power under a large field-of-view, we demonstrate the use of low power objective to resolve the entire architecture of large-scale neonatal mouse hearts, revealing the helical orientation of individual myocardial fibers. Therefore, our c-LSFM imaging approach provides multi-scale visualization of architecture and function to drive cardiovascular research with translational implication in congenital heart diseases.
Highlights
Micro–magnetic resonance imaging (MRI) are limited from tracking the fluorescently labeled live zebrafish and neonatal hearts for cardiac development, injury and repair
With its numerous structure variants, including Selective Plane Illumination Microscopy (SPIM), Multidirectional SPIM (m-SPIM), Digitally Scanned Light Sheet Microscopy (DSLM), Objective-Coupled Planar Microscopy (OCPI), Oblique Plane Microscope (OPM), and Bessel-Beam-based Light Sheet Microscopy, being widely developed in recent years, Light Sheet Fluorescence Microscopy (LSFM) is on the verge of becoming a predominant visualization technique for a broad range of life science research[12,13,14,15,16]
We have built on the established SPIM principle for tunable cardiac LSFM (c-LSFM) imaging to uncover cardiac architecture, development and remodeling otherwise challenging with existing imaging modalities
Summary
Micro–MRI are limited from tracking the fluorescently labeled live zebrafish and neonatal hearts for cardiac development, injury and repair In this context, Light Sheet Fluorescence Microscopy is an emerging imaging modality for multi-dimensional, -scale, and -channel visualization of cardiac architecture and physiology accompanied by rapid and precise tracking of multi-fluorescently labeled intracellular, cellular or tissue components from several microns to millimeters[9,10,11]. We further achieved cellular resolution to uncover the helical orientation of individual myocardial fibers, as well as the bi- and tri-cuspid valves, muscular ridges and trabecular network In this context, we have built on the established SPIM principle for tunable c-LSFM imaging to uncover cardiac architecture, development and remodeling otherwise challenging with existing imaging modalities. We introduced a widely tunable c-LSFM system for high-speed, large-FOV, and long-term multi-scale imaging ranging from zebrafish model to the higher vertebrate with translational implication to studying congenital heart diseases in the mouse models
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