To observe and analysis ultra-microscopically barley aleurone cell surface, atomic force microscope (AFM) was used. Seed coat of early maturing germplasm, eam9, was dehulled and scanned by non-contact mode. We have obtained the high resolution topographic 3-dimensional image of barley aleurone layer with high resolution. These images showed the membrane proteins in barley aleurone cell. One channel protein and numerous peripheral or integral proteins were detected in a area of 100 $mu extrm{m}^2$. Furthermore, we found that their widths were ranged from 50 to 750nm and lengths from 0 to 66 $mu extrm{m}$. The thickness of aleurone layer was measured by scanning electron microscope. The thickness at early developmental stage was about 16 and then the aleurone cell enlarged upto 57 $mu extrm{m}$${mu}{ extrm}{m}$ at least until 42 days after anthesis. In this study, we firstly reported on the ultrastructural AFM analysis of living aleurone cell as a biological specimen. It was clearly suggested that AFM will become an powerful tool for probing both the structural properties of biological samples.