A line-scanning ophthalmoscope (LSO) is a retinal imaging technique that has the characteristics of high imaging resolution, wide field of view, and high imaging speed. However, the highspeed imaging with rather short exposure time inevitably reduces the signal intensity, and many factors, such as speckle noise and intraocular scatter, further degrade the signal-to-noise ratio (SNR) of retinal images. To effectively improve the image quality without increasing the LSO system's complexity, the postprocessing method of image super-resolution (SR) is adopted. In this paper, we propose a learning-based multi-frame retinal image SR method that directly learns an end-to-end mapping from low-resolution (LR) image sequences to high-resolution (HR) images. This network was validated on down-sampled and real LSO image sequences. We evaluated the method on a down-sampled dataset with the metrics of peak signalto-noise ratio (PSNR), structural similarity (SSIM), and perceptual distance. Moreover, the power spectra and full width at half maximum (FWHM) were used as the no-reference image quality assessment (NR-IQA) algorithms to evaluate the reconstruction results of the real LSO image sequences. The experimental results indicate that the proposed method can significantly enhance the SNR of LSO images and efficiently improve the resolution of LSO retinal images, which has great practical significance for clinical diagnosis and analysis.INDEX TERMS Line-scanning ophthalmoscope, retinal images, multi-frame image super-resolution, learning-based.