Abstract: Diagnostic vitrectomy is a procedure that can obtain vitreous, retina or choroid specimens from patients with intraocular inflammation or malignant tumors of obscured etiology or irresponsive to treatments. An accurate diagnosis could be obtained from vitrectomy specimens by using related laboratory diagnostic technologies. Diagnostic vitrecomy with micro-incision and the developed laboratory diagnostic technology increase the specificity and sensitivity of disease diagnosing. The review summarized the surgical indications of diagnostic vitrectomy, specimen collection and process, methods of laboratory diagnostic technology, and the development of diagnostic vitrectomy.