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The CS/PVA/Fe3O4 nanocomposite membranes with chainlike arrangement of Fe3O4 nanoparticles are prepared by a magnetic-field-assisted solution casting method. The aim of this work is to investigate the relationship between the microstructure of the magnetic anisotropic CS/PVA/Fe3O4 membrane and the evolved macroscopic physicochemical property. With the same doping content, the relative crystallinity of CS/PVA/Fe3O4-M is lower than that of CS/PVA/Fe3O4. The Fourier transform infrared spectroscopy (FT-TR) measurements indicate that there is no chemical bonding between polymer molecule and Fe3O4 nanoparticle. The Fe3O4 nanoparticles in CS/PVA/Fe3O4 and CS/PVA/Fe3O4-M are wrapped by the chains of CS/PVA, which is also confirmed by scanning electron microscopy (SEM) and x-ray diffraction (XRD) analysis. The saturation magnetization value of CS/PVA/Fe3O4-M obviously increases compared with that of non-magnetic aligned membrane, meanwhile the transmittance decreases in the UV-visible region. The o-Ps lifetime distribution provides information about the free-volume nanoholes present in the amorphous region. It is suggested that the microstructure of CS/PVA/Fe3O4 membrane can be modified in its curing process under a magnetic field, which could affect the magnetic properties and the transmittance of nanocomposite membrane. In brief, a full understanding of the relationship between the microstructure and the macroscopic property of CS/PVA/Fe3O4 nanocomposite plays a vital role in exploring and designing the novel multifunctional materials. [ABSTRACT FROM AUTHOR]