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Adhered bacteria onto different surfaces cause infection that affects our health and environments. The understanding of the bacterial adhesive strength is crucial for better control and safe manufacturing in order to design adhesion resistant materials. The current evaluation methods lack precision and are often time consuming. In the present research, we developed a fluid-shear method to quantitatively evaluate bacterial adhesive strength on glass substrates. The glass was chosen based on its abundance in household, industrial, and medical environments. The fluid shear stress applied by a rheometer ranged from 0 to 3 Pa and the average surface roughness (Ra) of glass ranged from 1 to 23 nm. Bacterial adhesive stress was calculated based on the measurement of the critical radius. It was also found that the adhesive strength decreased with the increase of surface roughness, while the number of adhered bacteria increased when the surface become rougher. The fluid-shear method was proven to be effective in measure bacterial adhesion on a surface. [ABSTRACT FROM AUTHOR]