We have designed, fabricated, and tested a novel three-dimensional (3-D) flexible microprobe used for recording the neural signals of lateral giant (LG) on the escape system of American crayfish. We report an electrostatic actuation process to fold the planar probes to be the arbitrary orientations of 3-D probes for neuroscience application. The batch assembly method based on electrostatic force techniques gave more simple fabrication compared to others. A flexible probe could reduce both the chronic inflammation response and material fracture when animal breathes or moves. Furthermore, the cortex corresponds to hypothetical cortical modules with mostly vertically organized layers of neurons. Therefore, the 3-D flexible probe suits to understand how the cooperative activity for different layers of neurons. Advisedly, we present a novel fabrication for the 3-D flexible probe by using Parylene technology. The mechanical strength of the neural probe is strong enough to penetrate into a biogel. At the end, the flexible probe was used to record neural signals of the LG cell from American crayfish.