Abstract: Different methods were applied to specimen preparation of plant pollens for SEM observation. The results of comparative observations indicated that the micro-structure of plant pollen morphology was greatly influenced by specimen preparation. Three categories of situations occurred when the specimen of plant pollens were prepared by common preparation methods. The pollen appearance and ultrastructure of Pleioblastus amarus and the species in genus of Populus changed seriously because the pollen cell wall of these plant species were quite thin and the water content was high. With thicker cell wall and lower water content, the appearance of Hemerocallis fulva pollens did not change much, while the ultrastructure was distorted by shrinking. The pollen appearance and ultrastructure of Liriodendron tulipifera maintained well for its thick cell wall and low water content. It was pointed out that appropriate preparation method should be applied according to the characteristics of the pollens. For the pollens with thin cell wall and high water content, some measures should be taken to fix the specimen and to eliminate the influence of surface tension in order to show the appearance and ultrastructure characteristics more objectively.