Insects are increasingly common in households, zoological collections, research, and food industries. Increased knowledge of insect health parameters is necessary to ensure proper welfare. The study goal was to develop a hemolymph collection technique and report hemolymph serum biochemical parameters for the thorny devil stick insect (Eurycantha calcarata). Clinically healthy adult stick insects (5 males and 14 females, 15.1–24.7 g) were enrolled. Four collection techniques were evaluated. Hemolymph collection was unsuccessful in males, but was successful in females by using a single technique. The insect was manually restrained in an elevated position and an 18-ga × 2.54-cm needle was used to puncture the membrane just caudal to the third pair of legs. With the puncture site directed ventrally, ≤1 ml of hemolymph was collected via gravity and allowed to visibly clot. The sample was then centrifuged, and the serum was separated. Serum samples were individually analyzed (Avian/Reptilian Profile Plus, VetScan VS2, Abaxis, Inc, Union City, CA 94587, USA). Fourteen samples (0.2–1.0 ml) were collected from 14 females. Median (minimum–maximum) parameters included the following: aspartate aminotransferase 12 (0–45) U/L, creatinine kinase 25 (0–76) U/L, uric acid 7.5 (3.1–13.7) mg/dl, glucose 12 (8–22) mg/dl, calcium 18.6 (17.2–19.4) mg/dl, phosphorus 15.0 (n = 1) or >30.0 (n = 13) mg/dL, total protein 2.7 (1.6–2.9) g/dL, albumin 0.9 (0.2–1.2) g/dL, globulin 1.7 (1.6–1.8) g/dL (n = 6) or not quantified (n = 8), potassium 10.6 (9.0–11.8) mmol/L, sodium < 100 mmol/L, and bile acids 0 lmol/L. This is the first report of biochemistry parameters in clinically healthy female stick insects. Larger sample sizes are needed to establish statistically valid reference ranges. Hemolymph collection techniques for male stick insects warrant further investigation.