The three-dimensional (3D) spheroid invasion assay can provide a rapid, automatable invasion system to determine cell invasion such as tumor cell. This method is also highly reproducible and has many other advantages than other invasion assays. During the process of three-dimensional (3D) spheroid invasion assay, spheroids will be formatted in a 96-well plate and each well will format one spheroid. A semi-solid matrix is generated by the BMM added directly to each well into which tumor cells invade from the spheroid body.