Surface Enhanced Raman Scattering (SERS) enhancement becomes higher with decreasing of interparticle gaps between silver-nanoparticles on Anodic Aluminum Oxide (AAO) substrates. The Raman imaging system provides spatial distribution which the Raman spectrum system doesn’t have. The SERS images show that the strong enhancement comes mostly from the appearance of hot-spots and these hot-spots are random distributed. The temporal response of hot-spots shows that there are two modes, spike and step, of hot-spots. The Raman imaging system provides a time-saving method to get the spatial distribution of Raman signal, which facilitates the use for biological sensing applications.