Dynamic control strategies to adjust vacuum (flow-controlled vacuum, FCV) and pulsation (flow-controlled pulsation, FCP) settings during a milking offer unique opportunities to meet individual cows’ physiological requirements for pre-milking stimulation and elicit their maximum milk ejection capacity. To date, the potential of these automated control strategies for optimizing the milk harvesting process in high-producing dairy cows with a thrice-daily milking schedule has not been investigated by rigorous methods. We lack objective and automated methods to efficiently identify cows with delayed milk ejection in need of an enhanced stimulation protocol, and to assign them in an automated manner. We propose to study the combined effect of FCV and FCP (FCVP) on milking performance and post-milking teat tissue condition in high-producing dairy cows milked three times per day in a rotary milking parlor. This controlled trial will be conducted at a 4,300-cow dairy with a thrice-daily milking schedule in New York State. Following a switch-back design, all cows in the milking herd will be assigned to treatment and control over a 12-week study with two 3-week treatment and control periods, respectively. Treatment will consist of FCVP and implemented at the cow/cluster level in conjunction with early milking unit attachment. During the control periods, cows will be milked with the conventional milking routine and machine settings that are currently employed on the dairy operation. Milking performance will be measured by analyzing data on milking characteristics (i.e., milking duration, milk yield, average milk flow rate, and peak milk flow rate) and milking irregularities (i.e., forced retract, kick-off, milking liner slip) obtained from electronic on-farm milk meters. Post-milking teat tissue condition (i.e., short-term and long-term changes) will be assessed at the end of each period through manual and visual assessment. We will use appropriate statistical methods to evaluate the effect of FCVP on each of the selected outcome variables. We expect that FCVP will elicit cows’ maximum milk ejection capacity through meeting their physiological needs for pre-milking stimulation and enhance milking performance while maintaining superior teat tissue condition.