A method of manufacturing the pixel structure is provided. The method includes forming a gate, a scan line connected to the gate, and at least one auxiliary pattern on a substrate. An insulating layer, a semiconductor layer, an ohmic contact layer, and a photoresist layer are formed in sequence. Afterwards, a single exposure and development is performed on the photoresist layer to form a first portion and a second portion. Next, the ohmic contact layer and the semiconductor layer which are not covered by the photoresist layer are removed to expose a part of the insulating layer. Next, the second portion of the photoresist layer is removed. Subsequently, a part of the thickness of the semiconductor layer not covered by the first portion is removed and the exposed insulating layer is removed, so as to form a channel layer and an insulating layer.