The present invention relates to a donor substrate and a method of manufacturing a light-emitting device. The donor substrate includes a reflective layer including an opening portion, a light absorption layer covering the opening portion of the reflective layer over the reflective layer, a heat insulating layer including an opening portion in a position overlapped with the opening portion of the reflective layer over the light absorption layer, and a material layer including a light-emitting material covering the opening portion of the heat insulating layer over the heat insulating layer. A target substrate and the donor substrate are disposed to face each other, and an EL layer is formed over the target substrate by performing light irradiation from a rear surface of the donor substrate.