Protocol for quantifying vertebral column morphology for the statistical analysis of scoliosis severity in zebrafish

Here, we present a protocol for quantifying zebrafish vertebral column morphology using micro-computed tomography (microCT) datasets for scoliosis researchers. We describe steps for orientation of skeletal and cranial microCT datasets in DataViewer, vertebral position measurements in ImageJ/Fiji, and analysis of vertebral position in R. The approach described here graphically depicts vertebral position while providing a means to quantify scoliosis severity and perform statistical analysis of differences in vertebral column morphology in zebrafish. For complete details on the use and execution of this protocol, please refer to Voigt et al.¹.