- 0.1% Tween PBS
- 3% paraformaldehyde
- Methanol (Stored at -20°C)
- Dissecting Dish
- Drawn Pipette
- Eppendorf tubes
- With the drawn pipette, remove as much of the solution as possible. Be careful not to disturb the dissected worms.
- Add 2 mL of 3% paraformaldehyde (PF) down the side of the dissection dish. Cover with parafilm, and leave in the dark for EXACTLY 1 HOUR.
- Add 1 mL of 0.1% Tween PBS down the side of the dissection dish. Swirl the dish gently to get the gonads in the middle of the plate.
- With a drawn pipette, gently collect the worms into 2 eppendorf tubes.
- Spin gently 1000 rpm for 1 minute in centrifuge.
- Wash with 0.01% Tween PBS.
- Add 1 mL of MeOH. Place in -20°C for 5 minutes.
- Wash 3x with 0.1% Tween PBS.
- If necessary, proceed to Step 6 in the DAPI staining protocol.