Accurate collection and preservation of vitreous and retina-related tissue specimens is the basis for clinical diagnosis and rigorous basic research. The clinical uses of vitreous specimens include microbial culture, cytological detection, detection of degenerative diseases, PCR analysis, and cytological detection of cell morphology. The experimental research uses include DNA gene analysis, protein quantitative analysis, metabolite examination, RNA content quantitative analysis, cytokine determination and so on. Retinal specimens collecting was mainly used for PCR analysis of retinal proliferative membrane, immunohistochemical staining, immunofluorescence examination, microvascular density evaluation, cell isolation and culture, etc. Understanding the collection of vitreoretinal surgical specimens and the application of relevant detection techniques and materials can provide a more comprehensive idea for the diagnosis of vitreoretinal diseases and a broader reference for the related basic research.