A continuous near-field electrospinning (NFES) process has been demonstrated to be able to achieve direct-write and well-aligned chitosan/poly(ethylene oxide) nanofibers. The ability to precisely control and deposit chitosan-based nanofibers in a direct-write manner is favorable in manipulating cells attachment and proliferation at a preferred position. Experimental results show that fiber diameters can be reliably controlled in the range of 265-1255 nm by adjusting various operating parameters of the NFES processes. These prescribed patterns of nanofibers exceed tens of centimeters long and complex configurations such as grid arrays and arc shapes are assembled at specified separations as small as 5 mu m. FTIR analysis reveals that NFES nanofibers have a similar morphology and composition as conventional electrospinning counterpart and constitute all components formerly present in the polymer solution. The versatile functionality to fabricate chitosan-based nanofibers with controllable size and directional alignment as well as highly ordered and customized patterns may represent an ideal candidate of a functional biomaterial and in tissue-engineering scaffolds that are predominantly representative of extracellular matrix (ECM).