Glomerular mesangial cells (MCs) are centrally located in the glomerulus. MCs control not only glomerular filtration, but also the response to local injury, including cell proliferation and basement membrane remodeling. Angiotensin II (Ang II) plays an important role in kidney function regulation, and participates in the progression of renal damage, as well as mesangial injury. However, studies on Ang II effects on MCs have used indirect methods, such as gene and protein expression after MC injury. In this study, we visually observed structural and mechanical changes to MC after Ang II treatment using atomic force microscopy (AFM). We obtained AFM topography and deflection images of live MCs, as well as fixed MCs in liquid, before and after Ang II treatment. Real-time imaging showed the dynamic movement of live MCs induced by Ang II. Changes in MC elastic property after Ang II treatment were measured using force-distance curves. AFM images of fixed and live MCs showed that cells contracted after Ang II exposure, with the nucleus height increasing within 20 min of Ang II stimulation. Force-distance analysis showed that Ang II caused MCs to stiffen (p < 0.0001). In conclusion, we demonstrated that AFM is an effective tool for real-time monitoring of live cell responses to drugs and stimuli.