protocol_376.txt

Schistocephalus solidus culturing
For each culture tube fill half way up with horse serum and then fill up with RPMI – leaving about 2 cm gap from the top.
Add 500µl of penicillium With a pair of blunt forceps gently open the top of dialysis membrane and place the worm into the membrane.
Screw the top on to the tube – and loosen slightly and place into the shaking 40°C water bath 
Check the worms each day – the worms may start to shed eggs after day 5 so can be collected after day 6 – if not collect eggs on day 7 – if the worms are left longer they may dissolve and the eggs will be difficult to collect cleanly
Remove the dialysis membrane from the culture tube and cut it open above any visible eggs, flush out onto a petri dish using ddH20.
Fill the petri dish to about ¼ full of ddH20
Under the stereo microscope with cold light source tilt the plate and the eggs should line up on the surface.
Collect the egg layer with a glass pipette and transfer to fresh petri dishes – up to 4 depending on the number of eggs.
Fill all the plates up to about half full with ddH20 including the original petri dish.
Label each plate with an identification number/name
Wrap each plate individually in foil and label the outside with the same identification plus your initials.
Place the wrapped plates in the 20°C incubator and leave in there for at least 21 days before checking for hatching