A microfluidic chip for culturing and real-time monitoring of multicellular tissues and use method thereof. The chip comprises a glass substrate layer, and a PDMS microchannel layer located on the glass substrate layer, wherein the glass substrate layer comprises a glass substrate, and a plurality of microelectrodes thereon; the PDMS microchannel comprises a plurality of independent microfluidic channels; the microelectrodes on the glass substrate are in one-to-one correspondence with the microfluidic channels in the PDMS microchannel layer; and the microelectrodes are electrically connected to an external circuit. The use method comprises: cell capture, cell or tissue culture, electrical impedance spectroscopy detection, and tissue release.