Long-term in toto cell tracking using lightsheet microscopy of the zebrafish tailbud.

Abstract

In toto light-sheet imaging allows the tracking of entire growing tissues with high spatial and temporal resolution for many hours. However, this technology requires a sample to be immobilised to ensure that the tissue of interest remains within the field of view throughout the image acquisition period. We have developed a method of mounting and image capture for long-term light-sheet imaging of a growing zebrafish tailbud from the 18 somite stage through to the end of somitogenesis. By tracking the global movement of the tailbud during image acquisition and feeding this back to the microscope stage, we are able to ensure that the growing tissue remains within the field of view throughout image acquisition. Here, we present three representative datasets of embryos in which all nuclei are labelled and tracked until the completion of somitogenesis.