There is provided a substrate separation device and method for separating a growth substrate from a laminate structure which includes a support substrate, a semiconductor layer, and the growth substrate. The device includes: a first base which is configured to hold the laminate structure thereon, and includes a first holding unit configured to hold the support substrate defining a bottom surface of the laminate structure and a heating unit configured to heat the laminate structure; and a second base including a second holding unit disposed above the first holding unit and configured to hold the growth substrate defining an upper surface of the laminate structure.