Travel demand has increased due to population growth, increase of vehicle ownership, and development patterns resulting in greater levels of congestion, pollution, and crash frequency. One approach to demand management is to increase the share of trips made by bicycles. With the increase in bicycling rates, there is a critical need for additional cycling infrastructure, which includes on and off-road bicycle lanes and paths, signs, markings, and signals. However, many of these infrastructure systems are implemented without detailed knowledge of bicyclist’s behavior and comfort while interacting with them. Therefore, if we understand factors that influence bicyclist’s behaviors and comfort on the roadway, planners and engineers will be better suited in implementing both bicycle technology and infrastructure. This study approached this challenge by evaluating a) bicyclist’s comprehension and preference to traffic control devices and b) bicyclist’s behavior and physiological responses to varying roadway conditions. Next, the research was interpreted for the purpose of improving practice within the transportation field. A survey questionnaire was used to evaluate bicyclists understanding and preferences of blue light detection feedback (BLDF) systems and bicycle signal countdown timers (BSCT). The results indicated that individuals understood and preferred the BLDF better with the additional novel signage that included text, symbols, and the blue dot. The study overwhelmingly showed that individuals “Strongly Agreed” that signage helped them understand the purpose of the BLDF, that they would support he implementation of the system, and that they felt better about waiting at an intersection with this system implemented. Individuals also generally understood the purpose of the BSCT, with the highest correct response from the numerical BSCT. Additionally, participants preferred the numerical BSCT, in comparison to the circular and vertical disappearing dot options. The Oregon State University (OSU) bicycle simulator was used in conjunction with a survey questionnaire to evaluate bicyclists’ galvanic skin response (GSR) responses, velocity, and lateral position to varying roadway conditions and bicycling infrastructure. The results showed that when individuals cycled within a bicycle lane, they had a GSR reading 1.25 peaks per min less than when cycling in a mixed traffic condition. In addition, when bicyclists rode in the bike lane, bicyclists GSR reading and velocity were not affected by variations in vehicular volume or speed. However, lateral position was affected by vehicular volume. When bicyclists were in mixed traffic conditions, the GSR reading was not affected by vehicle speed; however, it was affected by the vehicular volume. In mixed traffic conditions, none of the variables influenced bicyclist’s velocity. For the lateral position, only the vehicular volume had a significant affect. In summary, the recommendations from this work suggest a design for the BLDF system that will provide bicyclists with better understanding and comfortable at an intersection. Additionally, while preemption numerical countdown timers are not currently approved by MUTCD for vehicles or bicyclists, evidence suggests that a circular disappearing dots BSCT, was the preference of survey respondents. Based on the results of the simulator research, bicyclist’s stress response was not affected by the vehicular volume or the speed of vehicles while riding in a bicycle lane, which indicates bicyclists generally feel more comfortable while riding. Additionally, the vehicular speed did not play significant influence into bicyclists’ stress response or behavior; therefore, limiting the amount of traffic provided on the roadway can still make bicyclists feel less stress, even if a bike lane is not present. Therefore, recommendations for bicycle facilities should aim to provide striped bike lanes if possible or limit vehicular volumes on roadways where bicyclists operate in mixed traffic conditions.