Syzygium cumini (jamun) as a natural histological stain: A sustainable alternative to hematoxylin and eosin

Background: Histopathological staining is crucial for tissue diagnosis, with hematoxylin and eosin (H and E) being the standard. However, synthetic dyes pose environmental and health concerns. Syzygium cumini , an anthocyanin-rich fruit, presents a potential natural alternative. This study evaluates the staining efficacy of S. cumini extract compared to H and E. Aims: The aim of this study was to assess the staining efficiency, uniformity, and differentiation of S. cumini extract and its suitability as a histological stain. Settings and Design: An observational study was conducted on 60 archival formalin-fixed paraffin-embedded tissue samples, including normal and pathological oral tissues, decalcified bone, normal decalcified teeth, and exfoliative cytology smears. Materials and Methods: S. cumini dye was extracted using distilled water, 45% glacial acetic acid, and 70% ethanol. Soxhlet extraction enhanced staining efficiency. Staining was performed at 10, 15, 20, and 60 min. Three blinded oral pathologists evaluated cytoplasmic and nuclear staining, uniformity, intensity, and differentiation. Statistical Analysis Used: The Chi-square ( Χ 2 ) test was employed using SPSS software (IBM Corp., Version 27) to assess the statistical significance of differences in staining characteristics between H and E and S. cumini staining. P < 0.05 was considered statistically significant. Results: S. cumini staining was comparable to H and E across all tissue types, with no statistically significant differences ( P > 0.05). The 70% ethanol extract at 20 min provided optimal results, and Soxhlet extraction improved consistency. Conclusion: S. cumini extract is a viable, eco-friendly histological stain for cytoplasmic and nuclear structures. Further research is needed to determine its long-term stability and clinical applications.