zebrafish larva preparation, mounting, and light-sheet imaging for cardiac development studies

Analysis of Zebrafish Cardiac Contraction with Virtual Reality Interaction

The zebrafish (Danio rerio) is a widely used model organism for studying cardiac development, physiology, and repair due to its optical transparency, genetic tractability, and regenerative capacity1,2,3,4. Upon myocardial infarction, while structural and functional changes impact the cardiac ejection and hemodynamics, technical limitations continue to hinder the ability to investigate the dynamic process during cardiac regeneration with the high spatiotemporal resolution. For example, conventional imaging methods, such as confocal microscopy, have limitations in terms of imaging depth, temporal resolution, or phototoxicity for capturing the dynamic changes and assessing cardiac contractile function during multiple cardiac cycles5.
Light-sheet microscopy represents a state-of-the-art imaging method that successfully addresses these issues by quickly sweeping the laser across the heart&#39;s ventricle and atrium, achieving detailed images with enhanced spatiotemporal resolution and negligible photo-bleaching and photo-toxic effects6,7,8,9,10,11.
This protocol introduces a comprehensive imaging strategy that includes LSM system construction, 4D image reconstruction, 3D cell tracking, and interactive analysis to capture and analyze the dynamics of cardiomyocytes across the entire heart during multiple cardiac cycles12. The customized imaging system and computational methodology allow&#160;one to track the myocardial microstructure and contractile function at the single-cell level in transgenic Tg(myl7:nucGFP) zebrafish larvae. Furthermore, small molecule compounds were delivered into the embryos using microinjection to assess drug-induced cardiac injury and subsequent regeneration. This holistic strategy provides an entry point to in vivo investigate structural, functional, and mechanical properties of myocardium at the single-cell level during cardiac development and regeneration.

Author Spotlight: High-Resolution 4D Light-Sheet Imaging and Virtual Reality in Zebrafish for Single-Cell Analysis of Heart Function

4D Light-sheet Imaging of Zebrafish Cardiac Contraction

Our research uses 4D light-sheet imaging to explore cardiac contractile function in zebrafish, aiming to understand the dynamics of cardiac contraction at the single-cell level. One of the foremost experimental challenges is achieving high spatial terminal resolution, dynamic imaging of cardiac function while observing natural heart rhythms. Our study fills the gap in analyzing intricate 4D cardiac dynamic at single-cell resolution.We utilize GPU-based parallel computation, virtual reality, and advance cell tracking to interpret light-sheet imaging data. Our novel virtual reality platform offers an interactive approach to assess regional cardiac contractions, providing manipulative functionalities for more in-depth analysis. By facilitating user-directed analysis of cardiac contraction, our protocol could lead to an enhanced understanding of cardiac development and disease.

Zebrafish Larva Preparation, Mounting, and Light-Sheet Imaging for Cardiac Development Studies

Research

JoVE Journal

Bioengineering

Zebrafish is an intriguing model organism known for its remarkable cardiac regeneration capacity. Studying the contracting heart in vivo is essential for ...

4D Imaging Reconstruction of Zebrafish Larva, 3D Cell Segmentation, and Cardiac Contractility Analysis in Virtual Reality