Making In Situ Baskets
Step 1 Cut eppendorf tubes
Using a new razor blade (for a few baskets) or a scalpel blade on a handle (for many baskets), remove the bottom of the tube, as well as the cap, flush with the tube. You can heat the scalpel blade by passing it through a Bunsen burner flame a few times. This is make cutting the plastic easier. Be careful as the blade and the handle near the blade will become very hot.
Cut the bottom of the tube as straight as possible. If the bottom is cut at a slant, the embryos will collect on one side of the basket and stick together. You can carefully trim the bottom after the initial cut if needed.
Using a variety of colored tubes is useful for sample identification during in situ protocol.
Step 2 Cut mesh into thin strips
Mesh: Sefar NITEX Nylon Lab Pak
Item #: 03-110/47
Mesh is available on Amazon.com:
Step 3 Heat hot plate and melt mesh onto tube
Heat the hot plate to approximately 106 – 165 C. Be careful of your hands and clothing/sleeves near the plate - it is very HOT!
Press the tube onto the fabric on the hot plate. You may need to use a little pressure, but keep watching because once the plastic starts melting, it can melt too much and cover too much of the bottom of the mesh.
Step 4 Trim baskets
Remove the basket from the mesh strip with scissors before proceeding to the next basket. Just cut a rough square; you will trim the basket more neatly later.
After all the baskets have been covered with mesh, go back to each one and trim the mesh flush to the plastic with sharp scissors or a razor blade.