Proteome research has become a new hot spot in the post-genome era. High-resolution two-dimensional gel electrophoresis (2-DE), which provides the most comprehensive analysis system of the whole proteome, was highly improved in recent years. With the development of computer techniques, the powerful and user-friendly image analysis systems appeared to help high-throughput, large-scale proteomic studies. Using new generation two-dimensional image analysis software, ImageMaster 2D Elite, the 2D gels of proteins extracted from cultured Schwann’s cells were processed. The analysis procedure, including image acquirement, spot detection, match, background subtraction, pI/M_r calibration, analysis results report and database query, were reported and discussed.