The Heihe River Basin is the second largest inland river basin in Northwest China and it is also a hotspot in arid hydrology, water resources and other aspects of researches in cold regions. In addition, the Heihe River Basin has complete landscape, moderate watershed size, and typical social ecological environmental problems. So far, there has been no detailed assessment of glaciers change information of the whole river basin. Therefore, it is urgent to conduct studies on the assessment of glacial change in this region. Landsat TM/ETM+ remote sensing images, digital elevation model data and 1:50,000 topographic map data were used in this research. Through integrated computer automatic interpretation and visual interpretation methods, the object-oriented image feature extraction method was applied to extract glacier information. Glaciers change data were derived from spatial analysis, and the glacier variation and its response to climate change in the period 1956/63-2007/11 were also analyzed. The results are shown in the following. (1) In the period 1956/63-2007/11, the Heihe River Basin's glaciers had an evident retreat trend, the total area of glaciers decreased from 361.69 km~2 to 231.17 km~2, at a rate of 36.08%, and average single glacier area decreased by 0.14 km~2; the total number of glaciers decreased from 967 to 800. (2) Glaciers in this basin are mainly distributed at elevations of 4300-4400 m, 4400-4500 m and 4500-4600 m; there are significant differences in glacial distribution and glacial change. (3) Compared with other western mountain glaciers, glaciers retreat in the Heihe River Basin had a higher rate. (4) By analyzing the meteorological data at six weather stations from 1960 to 2010, it is found that the mean annual temperature increased significantly and the annual precipitation also showed an increasing trend. It is concluded that glacier shrinkage is closely related with temperature rising. Besides, glacier melting caused by rising temperatures is greater than glacier mass supply by increased precipitation to some extent.