The forest-grassland ecosystem in the middle and upper reaches of the Yellow River has important ecological service functions and is the key ecological foundation for the sustainable development in the basin. Calculating the value of the forest-grassland ecosystem services functions in the middle and upper reaches of the Yellow River is of fundamental importance for grasping the ecological asset reserves and promoting high-quality development. The environmental conditions of the middle and upper reaches of the Yellow River are complex and diverse, and there are spatial differences in ecosystem service level. Therefore, it is an urgent problem to accurately assess the value of forest-grassland ecological services in this region from a micro-scale. Based on multi-source spatial data and spatial analysis method, high-resolution accounting and spatial pattern analysis of forest and grassland ecosystem services were carried out. This study integrated use of remote sensing data products, meteorological data, ecological statistical survey data and other multi-source data. A 1km×1km pixel scale forest and grass ecosystem adjustment service function value system was constructed by referring to the existing ecological service value evaluation norms and literatures. This system overcomes the limitations of traditional statistical monitoring data in terms of accuracy and pixel size, providing a basis for systematic and coordinated regional ecological governance. In 2015, the value of forest and grassland ecological service functions in the middle and upper reaches of the Yellow River basin was 1.90 trillion yuan, of which the value provided by the forest ecosystem was 1183.31 billion yuan, the value provided by grassland ecosystem was 716.46 billion yuan. The high-value areas of forest and grassland ecosystem service value per unit area were mountains with good hydrothermal conditions in Qinghai-Gansu, Shaanxi-Shanxi; The low-value continuous areas were widely distributed in the Loess Plateau, which was closely related to the natural conditions such as the forest depression, grassland coverage, regional precipitation and topography. The values of six forest ecosystem service functions were mainly distributed in the three regions of Qinghai-Gansu in the west, Ningxia-Inner Mongolia in the north-central, and Shaanxi-Shanxi in the east. The service function value of grassland ecosystem mainly presented a spatial pattern of "high in the south and low in the north". In the ecological protection and high-quality development of the Yellow River Basin, we should pay attention to the ecological service function as the basis and break through the boundary limits of the administrative region to establish the ecological product value realization mechanism and enhance the value weight of the ecological benefits.