Photobiomodulation therapy (PBMT) effects depend on the energy settings and laser penetration. We investigated the penetration time profiles of two different light therapy devices, at the dark and light skin regions in horses. Six light skin and six dark skin adult clinically healthy Arab and Quarter horses were used. A cutometer was used to measure the width of the skin fold from both sides of the cervical area, followed by three measurements of the thickness of the same skin fold by transversal and longitudinal ultrasonography (US). The depth of light penetration was compared based on the percentage of penetration versus power, between a portable PBMT device versus a class IV laser device. The laser mean power output was measured with an optical power meter system for 120 seconds after penetrating the skin. Skin width and laser penetration were compared among equipment by paired "t" test. There was no difference in the width of the skin fold between measurements acquired by the cutometer against either longitudinal or transversal US or between the US measurements at cervical versus metacarpus area. Light penetration was greater in both kinds of skins in the PBMT (0.01303 ± 0.00778) versus class IV laser (0.00122 ± SD 0.00070) (P < .001). The PBMT device provided a greater energy penetration than the class IV laser in unclipped light and dark skin, suggesting that the former may produce a better therapeutic effect. The color of the skin changes penetration profiles of PBMT.