A Hemocyte Disturbance Assay to Assess Hemocyte Re-Adhesion to Hematopoietic Pockets

To mechanically disturb resident hemocytes, select up to four to eight larvae and place them in a 2-milliliter microcentrifuge tube with approximately 0.5 grams of 600-micron glass beads and 0.5 milliliters of water. Vortex the tube by hand at speed 10 for one minute.

Retrieve the larvae from the glass beads by spilling the contents of the microcentrifuge tube into a Petri dish, and picking out the larvae with a paintbrush. For the recovery phase, place larvae in the previously prepared Petri dishes with a thin layer of fly food. Incubate the larvae for 45 minutes to allow the larvae to re-establish their hemocyte pattern. After the recovery period, continue with the bleed-scrape dissections.