Wed. Jan. 22, 2020 KCl in Geoduck Strip Spawn

Does priming with KCl matter?

This was a small scale experiment performed to determine if fertilization can be achieved without KCl priming.

Experimental setup:

  • ~10mL of eggs prior to KCl priming
  • ~10mL of eggs prior to KCl priming + 3 drops of sperm (fertilization started at 2:35pm)
  • ~300uL of KCl primed eggs + sperm mix (600mL eggs + 6mL sperm) (fertilization started at 2:54pm)

RESULTS:

  • No Sperm, No KCl (90 min post collection):

  • +Sperm, No KCl (90 min post collection, 70 min post-fertilization):

  • +Sperm, +KCl (60 min post-fertilization):

Incubated samples over night at 4C

  • No Sperm, No KCl (~21 hours post collection, 100uL drop on slide):

  • +Sperm, No KCl (~21 hours post collection, ~20.5 hours post fertilization, 100uL drop on slide):

  • +Sperm, +KCl (~20.5 hours post fertilization, ~50uL drop on slide because that’s all that was left):

Link to all 20200121 photos

Link to all 20200122 photos

CONCLUSIONS

  • It seems like no embryo development happened in the no Sperm, no KCl sample
  • It seems like some embryo development did happen in the Sperm, no KCl sample after letting the fertilization go overnight, but they look kind of weird. I’m not sure if algae is eating them?
  • It seems like some embryo development did happen in the Sperm, no KCl sample after letting the fertilization go overnight. Most of these eggs seemed to lose pigment (does that mean they are dead or dying?)

Will have to discuss results with group

Written on January 22, 2020