In this paper, we studied the lateral deformation of human red blood cells (RBCs) during lateral indentation by an optically trapped silica bead with a diameter of 4.5 µm (Bangs Laboratories, Inc. Fishers, IN, USA). The images were captured using a CCD camera and the Boltzmann statistics method was used for force calibration. Using the Hertz model, we calculated and compared the elastic stiffness resulting from the lateral force, showing that the differences are important and that the force should be considered. Besides the lateral component, this setup also allowed us to examine the lateral cell-bead interaction. The mean values of the cell shear stiffness measured during indentation were 3.37 ± 0.40 µN/m for biconcave RBCs, 3.48 ± 0.23 µN/m for spherical RBCs, and 3.80 ± 0.22 µN/m for crenelated RBCs, respectively. These results show that this approach can be used as a routine method for RBC study, because it enabled us to manipulate the cell without contact with the wall.