Abstract: Supporting structure for primary and tertiary mirror is one of the most challenging technical points during development of off-axis three-mirror anastigmatism (TMA) biological imaging system. In order to minish the shape error of the mirror and meet the stability of the supporting structure which was working, the mirror supporting structure was designed and optimized with finite element method. First, based on requirement of optical design, materials and supporting mode for the mirror and its supporting structure were decided. Then, a weight reduction structure of the mirror cell was designed which had three-point backside and six-point lateral support structure. The flexible hinge with circular and cantilever beam was made according to the optimized mathematical model. The influence of the parameters on the precision of mirror surface were analyzed under the load conditions of gravity and uniform temperature rise of 4 ℃. Then, the static analysis and thermal analysis for the mirror subassembly were performed. The Finite Element Analysis (FEA) indicated that the surface error (RMS) was 1.529 nm at self-weight. RMS value was 2.426 nm when the temperature rised 4 ℃. Finally, the surface figure test and the wavefront aberrations test were carried out by using Zygo interferometer. The test RMS value of the surface figure is 0.025 λ and the wavefront aberrations of the optical system is 0.102 λ (RMS). The results show that this supporting system could meet the technical indicator requirements of biological imaging system (RMS value of the shape error ≤λ/40, RMS value of the wavefront aberrations≤λ/10).