Quercetin has various biological activities, but its poor water solubility and stability limit its applications. In this study, β-cyclodextrin was used as the host and quercetin was encapsulated in its cavity to prepare an inclusion compound. Then, a nanofilm was formed using electrospinning. Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), scanning electron microscopy (SEM), and thermogravimetric differential scanning calorimetry (TG-DSC) were used to characterize the properties of the inclusion compound nanofilms. SEM images showed that the nanofilm prepared by optimizing the electrospinning process parameters had a good nanofiber structure. XRD, FTIR and TG/DSC characterization of the nanofilm showed that quercetin was encapsulated in the cavity of β-cyclodextrin and was present in the nanofilm. The quercetin was slowly released from the nanofilm and still had good bacteriostatic effects on Staphylococcus aureus and Escherichia coli, indicating that the process of embedding and electrospinning did not affect the antibacterial activity of quercetin.