Contrast-Enhanced X-Ray Micro-Computed Tomography as a Versatile Method for Anatomical Studies of Adult Zebrafish

Abstract

One attractive quality of zebrafish as a model organism for biological research is that transparency at early developmental stages allows the optical imaging of cellular and molecular events. However, this advantage cannot be applied to adult zebrafish. In this study, we explored the use of contrast-enhanced X-ray micro-computed tomography (microCT) on adult zebrafish in which the organism was stained with iodine, a simple and economical contrasting agent, after fixation. Tomographic reconstruction of the microCT data allowed the three-dimensional (3D) volumetric analyses of individual organs in adult zebrafish. Adipose tissues showed a higher affinity to iodine and were more strongly contrasted in microCT. As traditional histological techniques often involve dehydration steps that remove tissue lipids, iodine-contrasted microCT offers a convenient method for visualizing fat deposition in fish. Utilizing this advantage, we discovered a transient accumulation of lipids around the heart after ventricular amputation, suggesting a correlation between lipid distribution and heart regeneration. Taken together, microCT is a versatile technique that enables the 3D visualization of zebrafish organs, as well as other fish models, in their anatomical context. This simple method is a valuable new addition to the arsenal of techniques available to this model organism.