A novel method has been developed for the manufacture of a three-dimensional multi-electrode array (3D MEA), particularly, the shape of micro-tips can be varied by MEMS technology to construct different multi-electrode array. It improved the disadvantage of single electrode, which meant that it was available for multi-recording and analysis of a series of signals. Overcoming the difficulty of making interconnection line on 3D MEA chips via innovative fabrication process design. Using local isolation and removal of oxide films to forming electrodes and interconnections with oxide isolation covers. A 10 × 10 3D MEA with a distance of 100 μm between each other for further bio-neural signals was demonstrated, and a test on rabbit retina was also available. Besides, integrating pre-amplifier and built-in resistors with 3D MEA was brought out to hopefully increase the efficiency of sensing.