How Do Hornworms Respond with their Immune Cells to Tissue Damage? Modeling Immune Cell Regeneration in Invertebrates

1) Obtain a pharate fifth instar larva

2) Label a 1.5ml tube and add 1:100 PO inhibitor to prevent hemolymph oxidation

3) Hold larva between two fingers and snip tail with sharp dissection scissors

4) Bleed hemolymph freely into tube 

5) Store tubes on ice

6) To dissect imaginal cells from larvae, anesthetize animal on ice for 10 mins

7) Cut using scissors through the outer layer cuticle down the midline

8) Pin on four corners on the petri dish

9) Observe under dissection microscope and use forceps to identify and remove imaginal disc found on the upper right side close to brain.