We compared injury incidence and mechanisms among youth, high school (HS), and National Collegiate Athletic Association (NCAA) boys' and men's lacrosse athletes for the 2014-2015 to 2016-2017 lacrosse seasons. Multiple injury surveillance systems were used to capture 21 youth boys', 22 HS boys', and 20 NCAA men's lacrosse team-seasons of data during the 2014-2015 to 2016-2017 seasons. Athletic trainers reported game and practice injuries and athlete exposures (AEs). Injuries included those occurring during a game and/or practice and requiring evaluation from an athletic trainer and/or physician. Injury counts, rates per 1000 AEs, and injury rate ratios (IRRs) with 95% confidence intervals (CIs) were calculated. The injury rate in youth was higher than those reported in HS (10.3 vs 5.3 per 1000 AEs; IRR = 2.0; 95% CI: 1.6-2.4) and the NCAA (10.3 vs 4.7 per 1000 AEs; IRR = 2.2; 95% CI: 1.9-2.5). When considering time loss injuries only (restricted participation of ≥24 hours), the injury rate in youth was lower than those reported in HS (2.0 vs 2.9 per 1000 AEs; IRR = 0.7; 95% CI: 0.5-0.95) and the NCAA (2.0 vs 3.3 per 1000 AEs; IRR = 0.6; 95% CI: 0.4-0.8). The concussion rate in youth was higher than those in HS (0.7 vs 0.3 per 1000 AEs; IRR = 2.4, 95% CI: 1.1-5.2) and the NCAA (0.7 vs 0.3 per 1000 AEs; IRR = 2.1, 95% CI: 1.2-3.7). Injuries at the youth, HS, and NCAA levels were most commonly associated with stick contact, inflammatory conditions (including bursitis, tendonitis, and other unspecified inflammation), and noncontact mechanisms, respectively. Although the time loss injury rate was lowest in youth boys' lacrosse, the concussion rate was the highest. Injury prevention approaches should be specific to the mechanisms associated with each level of play (eg, equipment skill development in youth).