Single-cell RNA sequencing (scRNA-seq) is a powerful and rapidly developing emerging technology that allows complete analysis of cellular heterogeneity and provides bioinformation that is not available from holistic analysis compared to traditional bulk RNA sequencing (bulk RNA-seq).In recent years, the technique has been widely used in the characterization and detection of single cells and the study of biological mechanisms at the cellular level.Currently, the application of scRNA-seq in the field of ophthalmology has been reported.Gene expression at the single cell level has been studied in animal models, retinal organoids, primary human retina, retinal pigment epithelium (RPE), and choroidal visual systems.Similarly, scRNA-seq research has identified many ocular diseases under the background of the change of gene expression patterns, including how choroid blood vessel endothelial cell changes in age-related macular degeneration (AMD), the degradation of the function of cells in the retina of diabetic mice subtypes and myopic sclera fiber layer in the organization of various types of heterogeneous cells in mice, and so on.In addition, the scRNA-seq has identified the molecular, cellular and physiological characteristics of several eye tumors, including the cell origin and target molecules of retinoblastoma, the heterogeneity of uveal melanoma, and new therapeutic targets.This article mainly introduces the latest application of scRNA-seq technology in ophthalmology-related diseases to provide a reference for researchers, hoping that the technology can be extended to more related research in ophthalmology.