Hanie Seidli; Elham Shiri; Abbas Bakhtiari; Younes Mohammadi & Fatemeh Mirzaei
Histopathological analysis is critical for diagnosing disease and needs high-quality tissue preservation. Effective fixation is particularly crucial for ovarian tissue to maintain the detailed structures of the ovaries. This study systematically evaluated the effectiveness of three common fixatives, Neutral Buffered Formalin (NBF), Bouin’s solution, and Carnoy’s solution, on the morphological preservation of mouse ovarian tissue across fixation periods of 12, 24, and 48 hours. Ovarian samples from 27 adult NMRI female mice were fixed, processed, and stained with hematoxylin and eosin (H&E). Morphological preservation and stromal and follicular artifacts were estimated using a scoring system. Carnoy’s solution showed superior preservation at 12 hours, showing few artifacts and clearly distinct cellular components, although its quality notably deteriorated with prolonged fixation. Bouin’s solution delivered the best results at 24 hours, reaching a balance in cytoplasmic preservation and tissue staining, but it did not perform as well as Carnoy’s at 12 hours. NBF consistently produced poor results at all-time intervals, with a gradual decrease in artifacts over time, but inadequate preservation of follicles. Statistical assessment confirmed significant differences in the percentage of artifacts among the various fixatives and durations (p < 0.05). These results underscore the necessity of choosing fixatives according to the fixation duration: Carnoy’s solution for short-term (≤12 hours), Bouin’s for medium-term (24 hours), and NBF for prolonged preservation. This research offers essential insights for improving ovarian tissue fixation protocols, thereby increasing the accuracy of diagnostics and research in reproductive pathology.
KEY WORDS: Ovarian histology; Fixation artifacts; Carnoy’s solution; Bouin’s solution; Neutral Buffered Formalin; Follicular preservation.
SEIDLI, H.; SHIRI, E.; BAKHTIARI, A.; MOHAMMADI, Y. & MIRZAEI, F. Effect of different fixatives and duration of fixation on the histological integrity of mouse ovarian tissue. Int. J. Morphol., 44(1):350-357, 2026.
