Development of a Zebrafish Intubation Chamber for in vitro Longitudinal Imaging

Authors

• TJ Jones
• J Krynitsky
• P Gordon
• TJ Pohida
• K Tanner

Abstract

Zebrafish has been a model system for development and human pathologies. Despite technical advances, the full advantage of this model for real time visualization and multiplexed measurements for determination of biological mechanisms has not been fully realized. At the larval stages, fish are amenable to imaging as they are small with reasonable optical clarity and can be immobilized without deleterious effects. This imaging advantage is lost as the zebrafish develop into juvenile and adulthood. Simply, at these ages, zebrafish do not survive if the gills are not constantly stimulated by water flow. Thus, immobilization techniques such as a hydrogel or static microfluidics results in death of the animal. In addition, drug screens are routinely performed using larval stages and, while useful, these screens would be more applicable in the adult animal after development is complete. Here, we address both concerns by developing a system that provides the zebrafish with a steady flow of oxygenated system water at the physiological temperature of 28.5⁰C. Fish are appropriately sedated using an intubation system that delivers anesthesia. Oral gavage is also achieved by intubation thus mimicking what is achieved using mouse models instead of adding the drug to the fish water. Our zebrafish intubation chamber supports a single sedated zebrafish which then enables multi-hour imaging experiments on commercially available confocal systems such as a Zeiss LSM 780 confocal microscope.

Scientific Focus Area: Biomedical Engineering and Biophysics

This page was last updated on Tuesday, August 6, 2024