2020, Vol. 5 Issue 2, Part DECEMBER
Comparative studies of whole mount staining of bones and cartilage of selected bony fishes by Alizarin red S and Alcian blue
Author(s): Abhijit Dutta, Panchanan Das, Shristi Singh, Sonali Suprimayee Nayak, Reetika Sah, Asiya Anwar and Nilanjana Bhattacharyya Nath
Abstract: Comparative anatomical studies of bones and cartilages are rare in ichthyology as there is lack of reliable protocol for preparing the specimens that demonstrate all the bones and cartilages. The shapes and sizes of skeletal elements with their appropriate location were determined by whole mount skeletal staining. This technique is important for detecting changes in skeletal patterning. In the present study, five different fishes such as Amblypharyngodon mola, Macrognathus aureus, Puntius sarana, Sciades parkeri and Ambassis vachellii were treated with Alcian blue and Alizarin red S for the staining of cartilage and bone, respectively. Cartilages were stained blue because Alcian blue is a cationic dye which binds the sulphated GAGs and glycoproteins as cartilage contains higher concentration of GAGs than any other tissues. Bones were stained red because Alizarin red S is an anionic dye which binds with the calcium present in bones. Formalin was used as a fixative as it is a common fixative for museum specimen. This paper describes a procedure for rendering cartilage blue, bone red and soft tissue translucent or transparent in whole vertebrate specimen. This cleaning and staining procedure is thus readily applicable to comparative studies in anatomy, embryology, and systematic zoology.