Fri. Dec 7, 2018

Geoduck Broodstock Experiment

Respirometry

  • prepared 7 respirometry chambers
    • used 1/2 inch drill bit to drill out 3 holes/cover for cable glands. 9/16’’ would have worked better.
    • teflon taped around cable gland and fit into holes
    • used flex tape to seal the crack where the plug comes out
    • numbered each lid with sharpie and pump with paint pen

Geoduck hemolymph calcium assay

Question: Does calcium in hemolymph correlate with vitellogenin abundance/development stage?

prepared samples following Calcium assay protocol

  • standards A-H
  • for freezer samples, chipped out frozen sample with pipette tip and transferred to new tube to avoid thawing whole samples. Thawed chipped out samples on ice.
    • diluted hemolymph samples 1:1 in 1x Calcium assay buffer, 20 uL final volume
  • added 100uL of 1xPBS to ~20uL gonad tissue and homogenized with plastic pestle in blue 1.5mL tubes
    • spun at 12000rpm for 15 min at 4C and transferred supernatent to new tube
    • diluted 1:1 in 1x Calcium assay buffer (didn’t need to do this)

setting up plate reader

  • couldn’t figure out how to get the Seeb’s lab plate reader to read absorbance at 570nm. Asked Chris Monson, he said we may need a special filter. Contacted the sales rep he’s been dealing with to find out more. Contacted Krista, Mac, and Adam to see if we could use a plate reader at NOAA Montlake. They have one that could work but couldn’t use it until Monday. Contacted Emma and GS has a plate reader we could use Monday morning.

***saved samples in -80C (rack 5, column 3, row 2) and standards at 4C

Hemolymph Sampling Pilot

  • Chewy was found dead :( . No pics because it was not very pretty.
  • Star was still alive so I cleaned the tank and fed 5mL shellfish diet @1:45pm

After 3 hours, not much algae paste seemed cleared

Star’s siphon looked pretty catabolized and more blackened than usual, hope it’s not fungus.

Not sure if she’s going to make it, but we’ll see what happens next week



Written on December 7, 2018