Kaitlyn’s notebook: 20200121 Geoduck Strip spawn @ Pt. Whitney

Shelly, Brent and I helped Matt with a strip spawn yesterday!

  • Modified and tested 1000ul pipette tip and end of 50ml falcon tube for speculum
    • Inserted speculum into pedal gape
      • Used 18g needle to sample gonad
        • Put on slide and examined at 4X and 10X for sperm or egg

The modified falcon tube worked best because it was slightly larger and remained in place better when the geoduck contracted. Using a headlamp, we could actually see the gonad before taking a sample.

  • Tested 2 geoduck from each tank (1, 2, 4, and 5)
    • Found 3 ripe males and a ripe female
  • Shucked and removed gonad
  • Cross-hatched gonad; added egg and sperm into individual tripours
  • Rinsed eggs in 20um screen
    • lots of clogging from egg jelly –> used 2 screens
  • Primed eggs with ~4000 mL of 50mM KCL
  • Rinsed over 20um screen and added to 2L pitcher
    • Final volume of 1200ml; Cellometer estimated 4.2*10^6 eggs and microscope counts estimated 8.88*10^6 eggs

Checked on quality of sperm. 2/3 males had high quality sperm based on appearance and activity.

  • 600mL of eggs in 2 tripours
    • Added 6mL of sperm
  • Checked on scope for egg:sperm; over 30 egg to sperm egg-sperm
  • Added sperm to LRT (5000L tank) after 5 minutes.

The LRT was at 10C which may slow down the fertilization and development process. A sample after ~40 minutes showed some polar body formation.

 20200121_160841

Matt sent over a picture of polar bodies from the LRT later that day:
IMG_5611