Blood collection in chameleons is indicated in clinical cases and research studies and is commonly performed from the ventral tail vein. However, this venipuncture site can be associated with a greater degree of technical difficulty, adverse effects, and artifactual changes in the test results. The jugular vein is often suggested as a preferred alternative sampling site, but can also be technically challenging in smaller lizards. A transillumination method of the cervical region that reveals the jugular vein can be used for more accurate venipuncture, increased sampling success, as well as reduced handling time and soft tissue damage to adjacent anatomical structure. The technique described in this report can be used to collect blood from common chameleons (Chamaeleo chamaeleon).