In sports events, the rapid recovery after high-intensity training or sport competition performance is very important for athletes’ performance and health. The aim of this study is to evaluate the effect of laser acupuncture and electrical stimulation on the recovery from exercise fatigue, using mice with swimming fatigue as experimental model and the electromyography (EMG) and the Raman spectroscopy of blood as evaluation indicators. Root mean square (RMS) and mean power frequency (MPF) of EMG were analyzed after laser acupuncture and electrical stimulation. The amplitude frequency combined analysis (JASA) showed that the proportion of muscles in the fatigue recovery area of the control group, the laser acupuncture group, the multi-channel laser acupuncture group and the laser combined with electrical stimulation group were 34.78%, 39.13%, 39.13% and 43.48%, respectively. Raman spectroscopy of the mice blood during fatigue recovery showed there is a significant difference between the multi-channel laser acupuncture group and the laser combined with electric stimulation group compared with the recovery period and fatigue period (P<0.05) at the peak of 997cm− 1 and the laser combined electrical stimulation group had a statistical difference in the recovery period compared with the fatigue period (P<0.05) at the peak of 1561cm− 1. The results showed that laser acupuncture combined with electrical stimulation was beneficial to fatigue recovery in mice, and had the potential value in sports fatigue recovery.