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چکیده
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Objective: Tissue fixation is critical in histopathology for maintaining structural integrity. Despite its prevalence, the toxicity of 10% formalin has prompted the search for natural, non-toxic alternatives. Rosemary (Salvia rosmarinus), rich in phenolic and antioxidant compounds, presents a potential candidate for tissue preservation. This study evaluated the efficacy of methanolic rosemary extract as a fixative for sheep spleen tissue compared to standard formalin. Methods: Twenty-eight sheep spleen specimens were divided into four groups (n=7) and fixed in either methanolic rosemary extract or 10% formalin for 24 or 48 hours. Following fixation, tissues underwent routine processing and Hematoxylin and Eosin (H&E) staining. Histomorphometric analysis was performed to assess morphological preservation and cellular detail. Results: Histopathological evaluation revealed that rosemary extract provided acceptable structural preservation at both 24-hour and 48-hour intervals. The extract maintained the integrity of the capsule, white pulp, and red pulp, yielding results comparable to 10% formalin. Statistical analysis of cell counts showed no significant differences between the rosemary and formalin groups (P > 0.05). Notably, no evidence of autolysis or significant cellular degradation was observed in the rosemary-fixed samples. Conclusion: Methanolic rosemary extract serves as a viable, eco-friendly alternative for histological fixation, demonstrating comparable efficacy to formalin in preserving splenic architecture. While these results are promising for routine morphology, further research is required to evaluate its performance in immunohistochemistry and molecular applications, such as DNA/RNA preservation and PCR compatibility
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