A scroll is a unique form of display and collection for painting and calligraphy. However, creases on the surface of a scroll painting not only affect its appearance during exhibitions, but also reduce its lifetime. In our study, microstructures of scroll painting creases were analyzed by hyperfocal microscopy and scanning electron microscopy (SEM). We also discuss the mechanism of crease generation and propose a corresponding preventive measure. The results indicate that rolling-up with small winding diameter is one of the major causes of scroll painting creases. A smaller winding diameter makes that phenomenon more obvious. Generation of scroll painting creases and fiber breakage can be effectively reduced by addition of an axle tube, which increases the diameter of the winding rod. This proposed method can maintain traditional mounting forms and original appearances of paintings, and also preserve traditional charms of the art of painting and calligraphy.