Recently, minimally invasive surgery (MIS) has become apparent. Research has been performed on surgical robots needed in MIS. In the case of MIS, it is effective to express environmental impedance as numerical data in order to preserve it for establishing standard reference values of the conditions of internal organs. In this paper, we propose a novel method for the estimation of stiffness in real environments in which bilaterally controlled robots are needed. By using the proposed method, environmental stiffness can be estimated dynamically regardless of the initial position of slave system. The viability of the proposed method is confirmed from the experimental results.