We present here the first attempt to fabricate micro 3D structures on DMF chips for precise droplet splitting to achieve one drop multiple analysis. The structures were constructed by fabricating SU8 blades of different height on the dielectric layer. The droplet splitting is more consistent and controllable compared with the current 3-electrode splitting methods. With this new method, DNA multiplex detection was successfully performed in parallel with a single fluorescence channel. All these together had confirmed its potential for a wide range of multi-step biochemical applications including post-PCR molecular detection for point-of-care disease diagnostics.