This article presents the formation of a dextran nanofibrous layer by needleless electrospinning. Optimum process parameters such as polymer solution and addition (surfactant) concentration, voltage, distance, etc. were determined to obtain uniform and smooth dextran nanofibers. It was not possible to produce nanofibers from pure dextran/water solution. Instead, solution drops were deposited on the collector; therefore, anionic surfactant was added in various concentrations to start the nanofiber production. Also, the effects of surfactant concentration on the solution properties, spinnability and fiber properties were determined. Generally, uniform and fine nanofibers were obtained from the rod electrospinning method. The value of 2 wt% surfactant concentration was chosen as the optimum concentration to produce a dextran nanofibrous layer by roller electrospinning. According to the results, spinning performance was 0.6726 g/min per meter, average fiber diameter was 162 nm, diameter uniformity coefficient was 1.03 and the nonfibrous area was 0.5%. In conclusion, this methodology resulted in the production of good product properties such as good spinnability, fine and uniform nanofibers and high fiber density.