Staphylococcus aureus is the most frequent and major pathogen of mammary glands of dairy cows. There is increasing need to develop protective vaccine against S. aureus IMI. A good experimental infection model is required to evaluate vaccine efficacy. The intramammary infusion of S. aureus is a reliable method in terms of causing infection. However, intramammary infusion is unrealistic method in terms of mimicking natural IMI because it overwhelm the host immune mediated defenses since large number of bacteria were directly delivered into the intramammary area bypassing innate and acquired immune defenses at teat opening. Therefore, a challenge model that is similar to natural infection is required to evaluate protective effect of new vaccine against S. aureus mastitis. The objective of this study was to develop experimental S. aureus mastitis teat-dip infection model. A total of 8 dairy cows at early dry period were divided into group 1 (n = 5 cows) and group 2 (n = 3 cows). Cows in Group 1 were challenged with S. aureus strain SAUT1 by dipping all 4 teats in a suspension of S. aureus at a concentration of 10⁶ cfu/mL Tryptic soy broth (TSB) medium. Similarly, all 4 teats of cows in the group 2 (Control) were dipped in PBS. The challenge strain was grown to mid log phase in TSB at 37°C and cows were challenged after morning milking. Results showed that out of 5 S. aureus challenged cows 80% (n = 4 out of 5) and 20% (n = 1 out of 5) were infected sub-clinically and clinically respectively. At d 3 of challenge, at least one quarter of each cow was shedding S. aureus in milk resulting in 100% infection. At d 6 of challenge, 100% of challenged quarters were shedding S. aureus in milk. We also further evaluated the protective effects of experimental vaccine against S. aureus intramammary infection (IMI) using this model and found promising results. We concluded that teat dipping in the S. aureus bacterial suspension at cell density of 10⁶ cfu/mL of growth medium is good experimental intramammary infection model to induce S. aureus IMI.