Acta Vet. Brno 2024, 93: 321-330
The effect of aldehyde fixatives, alcohol-based fixatives, and fixatives combining denaturation and crosslinking on histomorphological, histochemical, and molecular genetic studies during short-term and long-term fixation: a comprehensive analysis
The choice of fixative solution has a significant impact on the quality of tissue processing and the results of ancillary pathological studies. The present study aimed to compare the effect of different fixatives and fixative times on routinely used histopathological staining methods and molecular genetic studies in porcine endometrium. Haematoxylin and eosin staining was used to evaluate cell morphology, while Gömöri and orcein staining were selected for histochemical studies; molecular genetic studies were represented by polymerase chain reaction. Tissue samples were fixed in formalin, Bouin’s fluid, methacarn, Greenfix, and Antigenfix for 1.5; 8.5; 15.5; 29.5; 64.5; 189.5; 249.5; 309.5; 369.5 days (only the period 1.5 days to 189.5 days was analyzed for histochemical studies). Formol proved to be the best for preserving cell morphology during both short-term and long-term fixation. Alcohol fixatives mainly affected the nuclear morphology, therefore they are not completely suitable for histomorphological analysis. The best preservation of nuclear morphology was achieved with Bouin’s fixative. For histochemical studies and short-term fixation, fixative solutions of all 3 groups (crosslinking, denaturing fixatives and fixatives combining both denaturing and cross-linking effects) were suitable. For long-term fixation, the alcohol fixative methacarn proved to be the best in histochemical studies. Finally, for molecular genetic studies, methacarn was the best fixative, while Bouin’s fluid was completely unsuitable. Formalin was confirmed as the most universal fixative overall. However, the selection of a suitable fixative should always be made depending on the specific pathological study and the fixation time.
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Funding
This study was supported by the project ITA of the University of Veterinary Sciences Brno (project no. 201011).
