Embryonic Lizard Skeletal Staining (Part 2)

After an additional 2 days in 0.1% Alizarin Red and 0.5% KOH, the skeletal prep is starting to look nice! The KOH further revealed the teeth and jawbone. You can see the calcification of the teeth below. (For reference see previous blog entries Hand Osteology, Advanced Lizard Embryo, Embryonic Lizard Staining Pt. 1) Red staining indicates calcium present in bone. Blue staining indicates cartilage (sulfated GAGs, which are negatively charged). Together, the bone and cartilage staining provide a clear picture of development in this late-stage embryonic lizard. 

Lizard embryo teeth and jaw

Below is shoulder and the vertebral column. Note that the soft tissues still have a substantial amount of Alcian blue, which has non-specifically binded. Next time, I will let the prep destain longer, to clear these tissues out more. 

Originally, I let the embryo sit at room temp for 2 days in Alcian blue (covered in foil). Then I proceeded to do an ethanol series from 100% - 75% - 50% - 25%. I let the embryo soak for a few hours each step. In hind sight, I probably should have let the embryo sit overnight in 100% ethanol. Extended time in ethanol will not damage the embryo.

Below are the lizard embryo's hand and digits. 

Below are the lower leg, feet and toes.

Anole lizard embryo toe pads

The lamella of the toes have an odd asymmetrical pattern of cartilage strips, where each toe is different.

Anole lizard embryo lamellae

Anolis carolinensis embryo lamellae

Anole lizard embryo toe pads

Anole embryo toes

Below, I have removed the outer scales and some of the lamella, revealing a "scaffold" like structure attached to the base of the toe. You can also see the bone beneathe, in pink.

Anole embryo toe pad

All embryos were treated ethically according to IUCAC protocol.

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