A manufacturing method of an image sensor of vertical type is provided that includes: forming an insulation layer with a metal wiring and a contact plug therein on a first substrate; bonding a second substrate having an image sensing unit over the insulation layer; forming a trench in the second substrate to divide the image sensing unit for each pixel; forming a PTI by gap-filling the trench with insulating material; forming a first material layer over the PTI, the image sensing unit, and the insulation layer; and forming a second material layer over the first material layer and performing a deuterium annealing process thereon. The crystal defects of the substrate generated when performing the trench etching on the donor substrate to define unit pixels are cured by performing the deuterium annealing process, making it possible to improve the sensitivity and illumination characteristics of the image sensor of vertical type.