The present invention provides an
evaporation mask plate, a manufacturing method thereof, an
OLED display substrate and an
evaporation method. The
evaporation mask plate comprises a body, evaporation opening areas formed in the body and a spacer arranged on the body. The spacer is at least arranged between adjacent evaporation opening areas and is used for evaporating organic light-emitting
layers of different colors. According to the technical scheme of the invention, during the evaporating process of organic light-emitting
layers on the
OLED display substrate, the evaporating materials of adjacent organic light-emitting
layers of different colors are blocked out by the spacer arranged on the evaporation
mask plate. Therefore, the
color mixing phenomenon of the organic light-emitting layers is avoided. As a result, no spacer is arranged on the
OLED display substrate, so that the process flow is reduced. The
process time is shortened. Moreover, the dimension of a pixel definition layer on the OLED display substrate is reduced, and the application amount of organic materials is reduced. At the same time, the resolution and the
aperture ratio of the OLED display substrate are improved.