Disclosed is a display substrate and a manufacturing method thereof and a display device. The display substrate includes an array substrate, an anode layer and a light-emitting layer which are sequentially stacked, and at least one film layer that is positioned on the light-emitting layer, wherein the at least one film layer includes a target film layer. The display substrate includes a first display area and a second display area which are adjacent to each other, wherein in the first display area, the target film layer has hollowed-out portions; and in the second display area, the target film layer is in a non-hollowed-out structure.