The present disclosure provides a display substrate, a manufacturing method thereof, and a display device. The display substrate includes a display area and a non-display area located at a periphery of the display area, wherein the display area includes a plurality of pixel opening areas and a pixel spacing area located between the pixel opening areas; the display substrate further includes: a first electrode, wherein at least part of the first electrode is located in the pixel spacing area, an orthographic projection of the first electrode on a substrate of the display substrate does not overlap an orthographic projection of the pixel opening area on the substrate; and a second electrode electrically connected to the first electrode and located in the non-display area.