The invention provides a micro-fluidic chip based on a droplet micro-fluidic technology and a detection method of the micro-fluidic chip. Cells can be monodispersed by adopting a spiral disperse phaseliquid inlet channel according to an inertial focusing principle, so that the cells are uniformly distributed in the channel from front to back, and single-cell droplets with high wrapping rate are favorably formed; besides, the disperse phase liquid inlet channel and a continuous phase liquid inlet channel are intersected in a cross shape to form a cross-shaped droplet generation channel, the flow ratio of two-phase liquid is convenient to adjust, the length and the distance of generated droplets are controlled, and the sizes of the droplets are more uniform and stable; and moreover, the micro-fluidic chip is provided with an upper layer and a lower layer, the upper layer is used for capturing high-flux single cells, and the lower layer is used for culturing the single cells and enriching secreta of the single cells, so that the single cells are cultured in droplets for a long time, and researches such as single cell in-situ culture, cell co-culture, drug screening, real-time and high-sensitivity detection of the secreta can be carried out, and the micro-fluidic chip is simple and flexible to operate, high in flux, pollution-free, short in consumed time, low in cost and wide in application range.