There are only few scientific publications dealing with the basic investigation of the effects of only one or two acupoints or comparing one single point with another single point, using different stimulation methods in the same persons. The aim of this needle-controlled, randomized crossover study was to investigate the neurovegetative parameters heart rate (HR) and heart rate variability (HRV) using two different acupoints, Baihui (GV20) and Neiguan (PC6), in separate sessions. We investigated 11 healthy volunteers (3 m, 8 f) with a mean age ± SD of 22.9 ± 2.8 years. The two acupoints were stimulated for 10 minutes each with manual needle acupuncture, red laser acupuncture (658 nm), and violet laser acupuncture (405 nm), in randomized order. Needle and red laser stimulation of the Baihui acupoint decreased HR significantly. Only violet laser stimulation at the Neiguan acupoint induced a significant increase of total HRV. Further studies using other neurovegetative parameters and more volunteers are necessary to confirm the preliminary results.