An appropriate calibration positioning method is imperative to examine localized tip on nanoscale patterns for scanning probe microscopy (SPM). This paper is to develop a new nanofabrication processes for AFM tip positioning with image stitching method in tip plowing technology. Moreover, this paper adjusts the set-point amplitude (A(sp)) to develop the tip plowing technology for fabricating nanopattern on 55 nm grating gage of a silicon substrate. The developed image stitching program is based on an iterative closet point (ICP) algorithm which has six degrees of freedom alignment. A closed-loop piezo motor is used to tip approach and plow in Z-axis. Experimental result of fabricating nanobagua on 55 nm grating of silicon substrate show that the developed positioning processes with image stitching method verify the feasibility of repeatability for the tip plowing technology successfully. This developed method can be further performed by a commercial atomic force microscope (AFM) with CAD/CAM. This technology can also be applied in dip pen nanolithography (DPN), SPM oxidation lithography and related fabrication technology with AFM tips.