In this study, chitosan (CTS) dissolved in acetic acid was electrospun together with aqueous poly(vinyl alcohol) solution (PVA) to prepare membranes (CTS/PVA) with nanofibers. The morphology of the membranes varied with the electrospinning conditions. High polymer concentration, high distance or high voltage was facilitative in obtaining a membrane with nanofibers. A CTS/PVA membrane with evenly distributed nanofibers (50 nm dia.) was obtained under the following conditions: 4.5 g CTS in 55 ml 27% acetic acid, 0.4 g PVA in 5 ml water (H2O), 0.1 g sodium chloride (NaCl), electrospinning voltage of 40 kV and distance of 25 cm from the injector to the collector. The water contact angle of CTS/PVA membranes increased after cross-linking with glutaraldehyde steam. Platelet adhesion measurement expressed that CTS/PVA with only nanofibers had no platelet adhesion, indicating good blood compatibility. Crosslinked CTS/PVA mats were stable while in contact with phosphate-buffered saline solution.