Development of a 3D Multi-Parameter Method to Evaluate Heart Looping and Chamber Volume in Zebrafish Embryos

Heart disease remains a leading cause of death worldwide. Some congenital cardiac malformations can result from defects in heart looping, which is a key process that ensures the correct alignment of the future chambers. The zebrafish is a widely used model for studying this essential developmental process; however, there is a lack of a standardized methodology to quantitatively characterize the heart looping process. Here, we present a standardized method for improving the accuracy when estimating the degree of heart looping using the zebrafish as a model. Our method relies on access to a lightsheet microscope, and 3D analysis software, and it provides a means to acquire multi-parameter measurements related to shape changes during the heart looping process. This simple and more accurate approach can be used in future studies, which attempt to quantify heart-looping defects as well as aid in the elucidation of components of the signal transduction pathways that regulate this process.