A manufacture method of an OLED substrate, the manufacture method comprises steps of: step 1, providing a substrate, and forming a plurality of anodes which are separately located on the substrate; depositing an inorganic film layer on the substrate and the plurality of anodes; step 2, employing a half tone mask to implement a patterning treatment to the inorganic film layer to obtain a pixel definition layer, a plurality of spacers located on the pixel definition layer, and a plurality of openings being located on the pixel definition layer and respectively corresponding to the plurality of anodes, and each opening exposes at least a portion of the corresponding anode to manufacture a first substrate; characterized in that the manufacture method further comprises steps of: step 3, coating a photoresist layer on the first substrate, and the photoresist layer covering the pixel definition layer, the plurality of spacers and anodes; cutting the first substrate coated with the photoresist layer to obtain a plurality of second substrates; step 4, stripping the photoresist layer on the second substrate to obtain a third substrate;and step 5, sequentially forming a Hole Injection Layer, a Hole Transporting Layer, an organic light emitting layer, an Electron Transport Layer, an Electron Injection Layer and a cathode from bottom to top in the plurality of openings on the third substrate to manufacture the OLED substrate. step 1, providing a substrate, and forming a plurality of anodes which are separately located on the substrate; depositing an inorganic film layer on the substrate and the plurality of anodes; step 2, employing a half tone mask to implement a patterning treatment to the inorganic film layer to obtain a pixel definition layer, a plurality of spacers located on the pixel definition layer, and a plurality of openings being located on the pixel definition layer and respectively corresponding to the plurality of anodes, and each opening exposes at least a portion of the corresponding anode to manufacture a first substrate; characterized in that the manufacture method further comprises steps of: step 3, coating a photoresist layer on the first substrate, and the photoresist layer covering the pixel definition layer, the plurality of spacers and anodes; cutting the first substrate coated with the photoresist layer to obtain a plurality of second substrates; step 4, stripping the photoresist layer on the second substrate to obtain a third substrate;and step 5, sequentially forming a Hole Injection Layer, a Hole Transporting Layer, an organic light emitting layer, an Electron Transport Layer, an Electron Injection Layer and a cathode from bottom to top in the plurality of openings on the third substrate to manufacture the OLED substrate. step 3, coating a photoresist layer on the first substrate, and the photoresist layer covering the pixel definition layer, the plurality of spacers and anodes; cutting the first substrate coated with the photoresist layer to obtain a plurality of second substrates; step 4, stripping the photoresist layer on the second substrate to obtain a third substrate;and step 5, sequentially forming a Hole Injection Layer, a Hole Transporting Layer, an organic light emitting layer, an Electron Transport Layer, an Electron Injection Layer and a cathode from bottom to top in the plurality of openings on the third substrate to manufacture the OLED substrate.