Disclosed is a micro-fluidic chip for high-throughput screening and high-throughput assay, in which its structure is improved, thereby enhancing the efficiency of high-throughput screening and high-throughput assay. The micro-fluidic chip includes a well for isolating a specimen. The well can be arranged in a one- or two-dimension. A specimen-isolating means is disposed above the well and is movable upwards and downwards. An opening and closing means for moving the specimen-isolating means upwards and downwards is disposed above the specimen-isolating means. An inlet for injection the specimen and an outlet for discharging an excess of the injected specimen are provided. A reagent-injecting passage for injecting a reagent and a reagent-discharging passage for discharging the reagent are also provided.