Liquid-liquid (L2) microlenses have great potential for various applications in imaging and detection systems. Traditional L2 microlenses are almost two-dimensional (2D) due to the modulation of flow rates in planer chips. Fundamental difficulties in effective application to cell imaging and analysis arise due to the limitations of 2D profiles. Herein, we demonstrate the feasible design of three-dimensional (3D) L2 biconvex lenses to detect flowing cells. Using the auxiliary curved microchannels, a 3D L2 lens is formed using Dean flow. The shape of the 3D biconvex lens and its focal length can be modulated by tuning the flow rates of the liquids. 3D light focusing was successfully achieved and the focal length could be modulated by around 435 μm, from 3554 μm to 3989 μm, in the experiment. The numerical aperture of the 3D L2 lens was also measured and its range was 0.175-0.198. Compared to a traditional objective lens with the same magnification (4×/0.1), the resolution of the 3D L2 biconvex lenses was improved 1.79-fold due to being completely immersed in liquid. Mouse myeloma cells sp2/0 and acute promyelocytic leukemia cells NB4 were imaged in the contrast experiments. The time response of experimental manipulation was about 2.7 ms. This 3D biconvex lens has great application prospects for cell imaging and analysis systems in lab-on-a-chip settings.