The present application provides an LTPS multilayered structure, which includes: a first stack layer having a reference pattern structure formed thereon and provided with uniformly distributed first references; and a second stack layer disposed on the first stack layer and having an alignment pattern structure formed thereon and provided with uniformly distributed second references each selectively aligning with one of the first references so that misalignment between the first stack layer and the second stack layer is precisely calculated by markings attached to each of the first references. The present further provides a method for measuring misalignment between a plurality of stack layers in the LTPS multilayered structure.