Three-dimensional (3D) spheroid assay provide a rapid and automate cell migration and cell invasion system in a highly reproducible, standardized methods. Multicellular spheroids are coated with cells and placed on the top of a conventional cell culture dish or in a 96 well-plate form as one spheroid per well. After the spheroids incubated with culture medium for a period of time and attachment of the spheroids to the plastic surface, the cells that have migrated from the spheroids can be monitored and cell movement can be measured by living cell microscope.