To build:
- cut a small gasket 3-5mm from thin plastic tubing, diameter equal to eppendorf.
- Fit this snuggly around a P1000 pipette tip so the resulting plastic and rubber-gasket fits very tightly in an eppendorf.
- Remove the inner plastic, put a small piece of nylon mesh over the rubber gasket, and fasten it in place by reinserting the inner plastic gasket.
- Cut the top of an eppendorf off just before it tapers. Insert plastic gasket with mesh membrane into this eppendorf with flat mesh end pointing towards the cut side.
- cut as second eppendorff tube about 1 cm above where it starts to taper.
- Cut a large gasket about 2 cm long from thick plastic tubing large enough to insert the cut eppendorfs, this holds the top of the first against the bottom of the second tube.
- Take a hyperdermic needle inserted through a P10 tip, use this to insert the tip through thick plastic tubing into the gap where the two eppendorffs meet. You may cut a small wedge at the edge of one to allow the ends to fit snuggly together around the P10.
- Remove the needle. This completes the assembly.
To use
- A P1000 tip inserted back first into the top of the eppendorf (with the cap) can be connected to a vacuum to drain all the fluid from the chamber. The mesh prevents the embryos from being aspirated.
- Adding fluid to the top chamber washes the embryos off the mesh and back into the bottom collection chamber. For thick fluids / detaching all embryos, apply the vacuum to the P10 tip while holding the assembly upright.
- To remove embryos, wash all embryos into bottom compartment using P10 vacuum assistance, then remove bottom compartment and pipette as normal.
- Be sure to rinse and wash the chamber in the appropriate solution prior to use.