Recent studies have shown that low-intensity laser therapy (LILT) enhances chronic wound healing, reduces pain, reduces inflammation, and improves post-operative rehabilitation. However, clinical outcomes in the veterinary use of LILT vary between different experimental studies. This is explained by improper laser parameter settings and limits of its penetration depth. This study aimed to investigate the penetration depth of 830 nm LILT on living dog tissue in different operating modes. This entailed continuous wave (CW) versus pulse wave (PW) and with contact versus non-contact techniques of the laser probe at different tissue-laser probe distances. The results can be applied for use in clinical practice.