Effects of the temperature of chitosan (CS) solutions as well as the temperature of the chamber on an electrospinning process were investigated. CS with a low molecular weight was dissolved in the solvent of trifluoroacetic acid/dichloromethane (70/30 v/v) at a concentration of 80 mg/mL for electrospinning. Both CS solution and chamber temperatures strongly affected the morphology of electrospun CS nanofibers. At the solution temperature and chamber temperature of 32°C, uniform CS nanofibers with an average diameter of 200 nm could be obtained. Although the chamber temperature is generally regarded as an unimportant parameter in the electrospinning of polymers, the experimental results demonstrated its critical effect on the electrospinning of CS.