2.3. Hemolymph collection
Oysterswere shucked open by severing the adductor muscle. Hemolymph was collected by penetrating the pericardial cavity located near the adductor muscle,
using a 21-gauge needle fitted to a 1 mL syringe.
Whole hemolymph was then transferred into sterilized 1.5 mL microcentrifuge tubes and stored in liquid nitrogen at −80 °C. Hemocyte lysates (HL) were produced by freeze-thawing the hemolymph
(−80 °C/room temperature) three times.
The lysed samples were centrifuged at 5000×g for 30 min (4 °C) to remove cellular debris.