Background: One of the reported pathways of cancer spread is the transcoelomic pathway, which is understood as the spread of cancer cells in the abdominal and thoracic cavities through interstitial fluid. It has been proven that the shear stresses caused by microfluidic currents on cancer tumors in the abdominal and thoracic cavities cause the detachment of cancer cells triggering transcoelomic metastasis; however, the magnitude of shear stresses has not yet been measured experimentally.Objectives: The objective of this study is to develop an experimental methodology using optical tweezers to approximate the shear stresses suffered by a nonporous, rigid artificial cancerous nodule model. Methods: Artificial cancerous nodule model was made by the agglomeration of 2 μm diameter polystyrene particles in a microfluidic platform. Optical tweezers were used as a velocimetry tool and shear stresses on the surface of the nodule model were approximated with the viscous shear stress equation. The results were verified with a numerical simulation performed in Ansys Fluent. Results: Shear stress originated by microflow over artificial cancerous nodule model were quantified both experimentally and numerically, showing good agreement between both methods. Such stress on the nodules' surface was much greater than that suffered by the wall on which the nodule model was located and dependent of the nodule model geometry. Although the experiment and simulation of this study were performed using a rigid and nonporous nodule model, the conclusion obtained about the increase of shear stresses applies to permeable, porous, and soft nodules as well, because the shear stresses are associated to the acceleration of the fluid originated by the reduction of the crosssectional area. Conclusions: Shear stress over artificial nodule model were successfully quantified using optical tweezer-based velocimetry technique and verified through numerical calculation. Advantages of experimental technique are: (1) it allows to control the position in a three-dimensional plane, allowing measurements in the vicinity of the analyzed surfaces, and (2) it is applicable for very low Reynolds number (R e « 1). On the other hand, as disadvantages: (1) it tends to be complicated to perform velocity measurements over obstacles and (2) it is limited in trapping distance.