Figure 5

Whole mount staining in early stages and applications of visualization techniques in adult zebrafish. Schematic representation of whole mount cleared and stained early stage zebrafish for cartilage with alcian blue, mineralized tissues (bone) with alizarin red and dual stained for both cartilage and mineralized tissues. Notice that only part of the skull, the basiventrals [for definition see Gadow and Abbott (243)] of the abdominal vertebrae and the fins endoskeleton are pre-formed in cartilage. Many bones in the skull and especially in the vertebral column are formed by direct intramembranous ossification. Images of adult skeletons taken by x-ray can be used to score for skeletal abnormalities, while microCT data can be used in an analysis program such as FishCuT to obtain quantitative data of bone measurements such as size, volume, thickness, and bone mineral density (80, 120). Bright field images or fluorescent images of whole mount cleared and stained zebrafish for mineralized tissues with alizarin red can be used to study skeletal abnormalities in detail. The three techniques are mostly used on euthanized and fixed specimens and thus can be applied on the same specimen sequentially. Moreover, the data procured by these visualization techniques can be integrated into a large data matrix and allows detailed phenotypic descriptions of zebrafish disease models.