A handling device is used for handling a substrate. The handling device includes a holder, a moving element, at least one first adhesive chuck, and at least one second adhesive chuck. The moving element is located adjacent to the holder for moving relative to the holder along a first axial direction. The first adhesive chuck is located on a first surface of the holder and the second adhesive chuck is located on a first surface of the moving element, in which the first adhesive chuck and the second adhesive chuck are used to be respectively separated from the substrate adhered thereon. In addition, another embodiment of the invention discloses a handing method of the handing device.