The invention provides a single-channel optical super-resolution imaging instrument which at least comprises an excitation light source module and an imaging, scanning and detecting module, wherein the excitation light source module comprises an optical laser, an acousto-optic modulator and an optical fiber coupler; the optical laser, the acousto-optic modulator and the optical fiber coupler are arranged in sequence; the optical axes of the optical laser, the acousto-optic modulator, a frequency scanning and selecting device and the optical fiber coupler are positioned on the same straight line; the imaging, scanning and detecting module comprises two lenses, a polarization beam splitter, a microscope objective, a sample stage and an imaging device; the optical axes of the lenses, the polarization beam splitter and the microscope objective are positioned on the same straight line and the optical axes of the imaging device and the polarization beam splitter are positioned on the same straight line. According to the invention, a single-channel frequency scanning, resonance excitation and polarization filtering combined method is adopted, so that the single-channel excitation super-resolution imaging of samples is realized. The single-channel optical super-resolution imaging instrument provided by the invention is simple in structure, obvious in effect and applied to the technical field of experiential and industrial super-resolution imaging.