Disclosed is a backside illuminated image sensor including a light receiving element formed in a first substrate, an interlayer insulation layer formed on the first substrate including the light receiving element, a via hole formed through the interlayer insulation layer and the first substrate while being spaced apart from the light receiving element, a spacer formed on an inner sidewall of the via hole, an alignment key to fill the via hole, interconnection layers formed on the interlayer insulation layer in a multilayer structure in which a backside of a lowermost layer of the interconnection layers is connected to the alignment key, a passivation layer covering the interconnection layers, a pad locally formed on a backside of the first substrate and connected to a backside of the alignment key, and a color filter and a microlens formed on the backside of the first substrate corresponding to the light receiving element.