Electrospun blend nanofibers were fabricated from chitosan (1,000 kDa, 80% DDA) and poly(ethylene oxide) (PEO; 900 kDa) at a ratio of 3:1 dispersed in 50% and 90% acetic acid. The influence of surfactants on the production of electrospun nanofibers was investigated by adding nonionic polyoxyethylene glycol dodecyl ether (Brij 35), anionic sodium dodecyl sulfate, or cationic dodecyl trimethyl ammonium bromide below, at, and above their specific critical micellar concentration to the polymer blend solution. Viscosity, conductivity, and surface tension of polymer solutions, as well as morphology and composition, of nanofibers containing surfactants were determined. Pure chitosan did not form fibers and was instead deposited as beads. Addition of PEO and an increasing concentration of surfactants induced spinnability and yielded larger fibers with diameters ranging from 10 to 240 nm. Surfactants affected morphology yielding needle-like, smooth, or beaded fibers. Compositional analysis revealed that nanofibers consisted of both polymers and surfactants with concentration of the constituents in nanofibers differing from that in polymer solutions. Results suggest that surfactants may modulate polymer–polymer interactions thus influencing the morphology and composition of deposited nanostructures.