Thurs. Nov 8, 2018

Geoduck Broodstock Experiment

Geoduck check-in and hemolymph sampling @ 3pm

  • both animals were still alive and OK
    • one animal had a starfish attached to back end
    • one animal had got its syphon through the holes in the container and a crab and urchin were eating it
  • sampled “Star” one from wrong end, pulled out tea-colored fluid
    • stuck from correct end once, but at too steep an angle
    • stuck again at ~45 degree angle and in front of (anterior to) the hinge and collected about 1mL of hemolymph
  • sampled “Chewy” one time in the correct area and got 1mL hemolymph on first try

  • visualized hemolymph under 10x and 40x and saw clusters of cells, but not as many cells as last week (no pics) - From Michael: “We definitely see a ton of variability day to day in hemocyte numbers. And the amount we saw here in you geoducks would have been a huge amount for a normal Mya arenaria. I am attaching those two photos. They were done on a slide with a convex surface, so a plain flat slide with a coverslip will have less volume and fewer cells visible. Also, this was after about 5-10 minutes, as I recall, and if the cells are allowed to settle for 30 or so, then most of them should be attached and spreading out.”

  • spun cells at 1000xg for 5 min at RT (because 4C centrifuge kept throwing Err 3). It seemed like most of the cells pelleted, although some may have still been in the lymph. Separated lymph from cells and froze at -80 C.

  • returned animals to original container by 4:30pm with small holes to protect them, and added air stone to it.

Lab meeting suggestions:

  • Try to compare pCO2 and TA to Sam’s and to other peoples chem data
  • Need to look up needle biopsy supplies
  • Setting up remote access to wireless router @ Pt. Whitney
    • put in access point mode so there isn’t communication confusion between both routers
  • determine pH to use: see Hollie’s paper where she used pH 6.8

Written on November 8, 2018