Shelly’s Notebook: 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): uc?export=view&id=11rXlDFYUJ0Wtmi4hdgStHdJtd0qPmK2J

uc?export=view&id=12OUDupaEHcN9OuWdPVnqkfFKwLI3dN3b

  • +Sperm, No KCl (90 min post collection, 70 min post-fertilization):
    uc?export=view&id=1wh8BwgPzqT5lXcmdh_YG6Uh_LWgIkhkI

uc?export=view&id=12Zm08ok7MktpYt7RwkQagk5iQtCa8rjg

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

uc?export=view&id=1_NIXDvOGT-MeqwgIgHY03thtMkdHUNvR

uc?export=view&id=1gWtmM42G76g2C5T12ZzOhEsuEQmzNtnw

Incubated samples over night at 4C

  • No Sperm, No KCl (~21 hours post collection, 100uL drop on slide):
    uc?export=view&id=1NVuM1TRBt3NN8YXC6ddKKxKosJcu8_M2

uc?export=view&id=1NrT0bzhlIQXBZofDyO9F0eScfYSm0OaP

uc?export=view&id=1CiEKIwxm9E2rv8e8UXKXDkxbmwUeGXcb

uc?export=view&id=1LP0C_a7X1dbe0jEeufk9biBtnaa4NdZM

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

uc?export=view&id=19VLvpMA9o6D6fTGYC8-Cr2c9O0NsjpUC

uc?export=view&id=1o71ve4tvb0dZZxFgvdz0rU8dDGWiXtsQ

uc?export=view&id=1C1SH-6qrEw7VXt3_V-YxRkHUr7uXAmiX

uc?export=view&id=1vdHHewW5LydPU_87jAUVqUvVGrrUMqnW

uc?export=view&id=1SFHEYbozigD-ONkya_bniPx8RdxcfHFZ

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

uc?export=view&id=18a5RacUvtiv_WveiQ1XYehuSrwmjOK9k uc?export=view&id=1RghO_psdRXQZCjRDNelt66LPT4zHltwm uc?export=view&id=1PQ878QMLDAj8jfHJW6Rz3Ui8H5WhXPp0 uc?export=view&id=1pegQe1O7SkKLD4RwOTaH5gjcC2N6td1C uc?export=view&id=1T5ztjoTNI409Z8jJAkazgvVnRk5BpuyI uc?export=view&id=1sw5qoO6m3mpOoZUdyNYe8YoRAuXjcHsj uc?export=view&id=1fp3je2-zsL-355HJQPfrps5rOo61YGsr

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

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