Purpose: Once cartilage has been stained, dissection of facial cartilage allows for efficient visualization and subsequent analysis of cranial cartilage formation.
Reagents and Equipment:
Hard surface for dissection – e.g. glass slide
Note: We first used agarose for the dissections but found it was too soft. We now dissect on a glass slide. Depending on the surface of the table, we place a white paper underneath the glass slide to provide contrast to visualize the stained cartilage.
Note: We previously made Tungsten needles to use for the dissections but found that Dumont #5 forceps were easier.
1. Place a stained embryo on a glass slide in 100% glycerol so that its ventral side is visible.
2. Hold the embryo firmly in place with the tweezers in your left hand posterior to its intestines. To avoid breaking the cartilage, grasp the adjacent tissue with the tweezers.
3. With the embryo in place, use the tweezers in your right hand to grab the tissue at the base of the intestines and pull it up, disconnecting the tissue from the cartilage.
4. Once the tissue on one side has been removed, the same steps can be applied to the other side in order to isolate the cartilage structure.
5. Better visualization of the cranial cartilage is possible by selectively removing additional tissues or adjacent structures.