Abstract: We observed pollen in the late development stages of fresh wild Arabidopsis thaliana by scanning electron microscope, using direct spraying of gold and the conventional biological electron microscope sample preparation method, and made comparative analyses. Arabidopsis pollen prepared according to the conventional biological electron microscope sample preparation method can maintain intact and clear outer wall structure under scanning electron microscopy. However, after direct spraying of gold, only mature pollen could be observed relatively clear outer wall structure, and no clear observation results could be obtained for pollen in other stages. Our results showed that the outer wall morphology of the mature pollen can be observed by direct gold spraying with fresh materials. This method is easy to operate and saves time. For the immature pollen, especially the pollen in the later stage of development, direct gold spraying will affect the pollen morphology, so we recommend using the conventional biological electron microscope sample preparation method to obtain the best shooting effect. The results of this study might provide some useful reference for researchers to observe the pollen outer wall structure in the late development stages of Arabidopsis thaliana by scanning electron microscope.