An image processing algorithm has been developed in order to analyze the nanofiber web images obtained from a high magnification microscope. It has been known that precise pore detection on thick webs is extremely difficult mainly due to lack of light uniformity, difficulty of fine focusing and translucency of nanofiber web. The pore detection algorithm developed has been found to show excellent performance in characterizing the porous structure, thus being a promising tool for on-line quality control system under mass production. Since the images obtained from an optical microscope represent only web surface, a scale factor has been introduced to estimate the web structure as a whole. Resulting web structures have been compared to those by mercury porosimetry, especially in pore size distribution. It has been shown that those two structures have a strong correlation, indicating that scaling of a single layer web structure can be an effective way of estimating the structure of thick fiber webs.