Pili or fimbriae are recognized as essential virulence determinants assembled on the bacterial surface. Gram-positive bacteria produce covalently linked pilus structures that are distinct from gram-negative counterparts. In this chapter, we describe three commonly used techniques to extract, detect, and visualize pili from gram-positive bacteria: (1) Western blot analysis, (2) Immuno-Electron Microscopy, and (3) Atomic Force Microscopy.