A separation device for a backlight source is provided. The separation device includes a machine table, at least one separation platform, where each separation platform is located above the machine table and secured at a fixed position relative to the machine table and is configured to support a display module and secure a backlight source of the display module thereon; a plurality of guide columns arranged perpendicular to the machine table, at least one adsorption mechanism arranged in a one-to-one correspondence with the at least one separation platform, where each adsorption mechanism is arranged at a side of the corresponding separation platform away from the machine table and configured to adsorb a display screen of the display module on the corresponding separation platform, and a driving unit configured to drive each adsorption mechanism to move in the lengthwise direction of the plurality of guide columns.