Biofilm as a stage in the bacterial life cycle has been the focus of many scientists for decades. This community composed by bacteria cells and extracellular compounds secreted by the cells play an important role in protecting the cells to survive from various threats, such as tough environments and antibiotics. There is an increasing need for bacterial control strategies with low environmental impact and can be produced in a sustainable manner. Surfactants are commonly used in cleaning products for its antibacterial ability, and biosurfactants which are produced by microorganisms from sustainable feedstock. Sophorolipids, as a biosurfactant, can disrupt mature Pseudomonas aeruginosa biofilm. By utilizing the microfluidic technology and microscopic imaging, we are able to create a micro-scale space with controlled flow of liquid which closely mimics the natural environment for bacterial activity. We evaluate how the sophorolipid affects P. aeruginosa initial attachment to the glass surface with continuous flow in microfluidic channels and glass bottom dishes. Through individual cell tracking and MATLAB processing code, we find that sophorolipid promotes the bacterial surface motility and inhibits micro-colony formation depending on the sophorolipid concentration, while sophorolipid shows no significant effect on the attached cell number reduction.