Objective: To investigate the utilization of environmentally-sustainable reagents in ophthalmic tissue pathology techniques. Methods: We prospectively collected 190 surgical specimens submitted to the pathology department of Zhongshan Ophthalmic Center between June 2023 and March 2024. Each specimen was divided into two sets, one treated with environmentally-sustainable reagents and the other with conventional reagents for fixation, dehydration, clearing, infiltration, staining, and mounting. The quality of histological sections from both groups was evaluated and compared. Results: Compared to the conventional reagent group, environmentally-sustainable reagent group showed good dehydration performance, moderately soft and hard tissue sections with intact cut surfaces and no cracks. H&E staining sections prepared with environmentally-sustainable reagents exhibited clear nuclear structures, distinct nuclear-cytoplasmic contrast, and clean backgrounds. The excellent section rates were 97.9% and 98.4% for the two, respectively. There was no significant difference in section quality between the two groups (Z = –0.645, P = 0.519). Tissue antigenicity was well-preserved during immunohistochemical procedures, with strong positive signals and accurate localization without background staining. Conclusions: The use of environmentally-sustainable reagents as a replacement for conventional reagents in the dehydration and staining of ophthalmic tissue yields satisfactory results, meeting the daily requirements for routine histological sectioning and staining. The entire process is xylene-free, environmentally-sustainable, cost-effective and worth being applied widely.
