Abstract: As congestion grows in metropolitan areas, agencies tend to utilize managed lanes on their freeway systems. Managed lanes have several forms and names, such as high-occupancy vehicle (HOV) lanes, high-occupancy toll (HOT) lanes, express lanes, and bus-only lanes. Although managed lanes have received significant attention as they increased the overall throughput and improved mobility without adding more lanes, little has been known about their operational capabilities. In addition, calibrating managed lane facilities can be challenging as they do not necessarily follow the same behavior with general purpose freeway lanes. This paper presents an operational analysis of two HOT lane segments located in South Florida. The sites are one-lane and two-lane segments separated by flexible pylons (FPs). The paper includes a macroscopic capacity analysis, and a microscopic calibration of the two sites using VISSIM microsimulation. The research findings assist in determining the capacity and speed-flow relationship of these segments, and also provide guidance for microsimulation model calibration for practitioners. The results of the study indicate that the percent drop in capacity for the one-lane FP site is 7.6% while the flow did not substantially change after the breakdown in the two-lane FP site. The research findings also include guidelines for simulating the breakdown events and calibrating one-lane and two-lane managed lane facilities in VISSIM microsimulation software. The Wiedemann car-following parameters (CC0 = 3.9 ft, CC1 = 1.9 s, CC2 = 26.25 ft, CC4 = −0.35, and CC5 = 0.35) provided the best fit for the one-lane FP site, while the combination (CC0 = 4.92 ft, CC1 = 1.9 s, CC2 = 39.37 ft, CC4 = −0.7, and CC5 = 0.7) parameters is recommended for the two-lane FP site.