Protein synthesis through translation is a fundamental and essential process for life. How translation begins, referred to initiation, dictates how the genetic code for each protein is read by the ribosome. Dysregulation of initiation has been implicated in various human disease states. I aim to understand the regulation behind initiation, specifically the process by which the ribosome and its cofactors locate the start site of translation. This process is dynamic, involving the rapid movements of the ribosome, posing a challenge to measure and quantify. To overcome these challenges, I plan to leverage a combination of single-molecule kinetics and mechanochemical measurements. With these approaches I will track the movement of the ribosome across an mRNA and uncover physical parameters of key factors in translation initiation.