Objective:

Bleed a few extra scallops (hatchery in the basement) from the F1 common garden culls ealier this year. Test SYBR green and JC-10 in hemolymph using the protocol written earlier this year with results summarized here

To do:

Again, the protocol has all the info one may need to go about this testing - check it out!

Beforehand… (day prior)

• filter seawater to 22um as your sheath (FSW)

• gather materials

  • syringe

  • needle(s)

  • ice and ice box

  • flow cytometry (‘FC’) tubes

• are all FC reagents full in the C6? If not, fill them and turn on the flow cytometry to ensure everything is in working order

• prep probes of interest

  • for example: JC-10, a 10xdilution of the stock solution allows for ease of pipetting to acheive 5uM in our hemolymph samples (review the hyperlink to the protocol above!)

view your sample animals

• below are some extra scallops in the hatchery from our experiment cull

gather materials

• assemble a mobile workspace akin to the photo below

Prep workspace (day of)

• gather all materials needed to bleed (syringes, needles, ice, ice box, FC tubes)

• prepare dark space (cooler, aluminum foil, box, blackout room, etc.)

• add 100ul 22um ice cold FSW to each tube for adding Hemolymph

• label tubes accordingly (scallop ID, treatment, FC probe, etc.)

sample your animals!

Results: