Electrospun nanofiber web has many potential applications due to its large specific area, very small pore size and high porosity. However, the mechanical properties of nanofiber web are very poor for use in textile application. To remedy this defect, the laminating process could accomplish in order to protect nanofiber web versus mechanical stresses. In this paper, direct tracking method as an image analysis based technique for measuring electrospun nanofiber diameter has been presented. The usefulness of the method for electrospun nanofiber diameter measurement is discussed. Such automated measurement of nanofiber diameter can be used to obtain better laminated webs.