A high throughput measurement method of human red blood cells (RBCs) deformability combined with optical tweezers technology and the microfluidic chip was proposed to accurately characterize the deformability of RBCs statistically. Firstly, the effective stretching deformation of RBCs was realized by the interaction of photo-trapping force and fluid viscous resistance. Secondly, the characteristic parameters before and after the deformation of the single cell were extracted through the image processing method to obtain the deformation index of area and circumference. Finally, statistical analysis was performed, and the average deformation index parameters (\begin{document}$ \overline {D{I_S}} $\end{document}, \begin{document}$ \overline {D{I_C}} $\end{document}) were used to characterize the deformability of RBCs. A high-throughput detection system was built, and the optimal experimental conditions were obtained through a large number of experiments. Three groups of samples with different deformability were used for statistical verification. The results showed that the smallest cell component \begin{document}$ \overline {D{I_S}} $\end{document} was 9.71%, and the detection flux of 8-channel structure was about 370 cells/min. High-throughput detection and characterization methods can effectively distinguish different deformed RBCs statistically, which provides a solution for high-throughput deformation analysis of other types of samples.