The aim of this study was to evaluate the effectiveness of pain modulation following Laser or LED phototherapies during the process of tooth separation. This was a longitudinal randomized controlled clinical trial in four observational times carried out in 60 patients (15 males, 45 females, average 24.1years old) who were randomly divided into three groups: G1 (LED, AsGaAl, λ850±10nm, 150mW, 17J/cm2, 57s per session), G2 (Laser, AsGaAl, λ780 nm, 70mW, 20.0J/cm2, 240s per session) and G3 (Non-irradiated Control). All patients were submitted to tooth separation using elastomeric separators. The pain level was measured by using a visual analogue scale (VAS) immediately after insertion (T1) of the elastic, at 48 (T2), 96 (T3) hours and 6days (T4). It was observed an increase of the pain on the Control group from T1 to T2, with statistical significance. Pain levels in the LED and Laser groups were always significantly lower (<0.001), except for T1. According with the results of the present study it may be concluded that, either LED or Laser phototherapies, were effective in reducing the pain level after dental separation process when compared to the control group.