Quartz crystal microbalance with dissipation (QCM-D) and dual polarization interferometry (DPI) were used to investigate fibrinogen (Fib) adsorption behavior on different surfaces by changing the pH value. Moreover, integrin adhesion to the adsorbed Fibs was studied using DPI. Qualitative and quantitative studies of platelet adhesion to the adsorbed Fibs were performed using scanning electron microscopy (SEM), confocal laser scanning microscope (CLSM), and released lactate dehydrogenase (LDH) assay. Experimental results indicated that the conformation and orientation of the absorbed Fibs depended on surface property and pH cycling. For the hydrophilic surface, Fibs adsorbed at pH 7.4 and presented a alpha C-hidden orientation. As a result, no integrin adhesion was observed, and a small number of platelets were adhered because the alpha C-domains were hidden under the Fib molecule. By changing the rinsing solution pH from 7.4 to 3.2 and then back to 7.4, the adsorbed Fib orientation became alpha C-exposed via the transformation of Fib conformation during pH cycling. Therefore, integrin adhesion was more likely to occur, and more platelets were adhered and activated. For the hydrophobic surface, the adsorbed Fibs became more spread and stretched due to the strong interaction between the Fibs and surface. alpha C-exposed orientation remained unchanged when the rinsing solution pH changed from 7.4 to 3.2 and then back to 7.4. Therefore, a large number of integrins and platelets were adhered to the adsorbed Fibs, and almost all of the adhered platelets were activated.