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INFORMATION FOR

Making Injection Dishes

Mesh coated injection dish

You will need the following supplies to make a mesh coated injection dish:

- 35 mm Falcon petri dish or any other polystyrene dish

-500(trop) or 800(laevis) micrometer polypropylene mesh. We use Spectrum Laboratories' Spectra/Mesh.

-methylene chloride

-fume hood

To make a dish, cut out an appropriately sized piece of mesh and place it in the Falcon dish. In a fume hood, draw up a small amount of methylene chloride. We use a 1 ml or 3 ml syringe, as it allows good control over the droplet size. Add small drops of methylene chloride to the center and around the sides of the mesh. After each drop, use a pencil eraser to press the mesh down at that spot for about 5 - 10 seconds. The methylene chloride will melt the polystyrene but not the polypropylene mesh. Try to keep the mesh flat as you go, working from inside out, or one side to the other, so there are not parts of the mesh bubbling upward. The embryos will fall through the mesh at these places. Also, avoid using too much methylene chloride, as it will melt the mesh to the point it won't hold the embryos. Be sure to work in a fume hood as the methylene chloride is very volatile.

Clear nail polish can also be used as an adhesive. Be sure to allow the polish to dry thoroughly before use.

Specifics for trop mesh from website:

SPECTRA/MESH PP 30CM/500UM 28498- PK3Spectra/Mesh

Polypropylene 30x30 350 cm

Mesh Opening (um): 500

Open Area (PK) (%): 39

Mesh Thickness (um): 610


Special thanks to L. Zimmerman, E. Amaya, P. Meade, N. Hirsch, and S. Borland (Indiana U-Axolotl Colony) for help with developing this protocol.