Cisternal puncture, although less frequently used since the advent of computed tomography and magnetic resonance imaging, is an important tool in the neurodiagnostic armamentarium. An anatomic study of cisternal puncture was conducted on fresh cadavers by direct visualization and fluoroscopic guidance of cisternal puncture. Anatomic and radiographic analyses demonstrated a dramatic dynamic anatomy of the cisternal space during cisternal puncture and indicated that “tenting” of the dura mater over the needle occurs reliably during cisternal puncture. Lateral fluoroscopy may decrease the risk of cisternal puncture by allowing visualization of needle depth in the cisternal space.