Potential of Green Betel Leaf (Piper betle L.) Extract as Natural Fixative in Histopathology

The use of 10% Neutral Buffered Formalin (NBF) as a standard fixative in histopathology has limitations due to its toxic and carcinogenic properties, thus requiring safer and environmentally friendly alternatives. Green betel leaf extract (Piper betle L.) contains bioactive compounds such as phenols, flavonoids, and tannins that have potential as natural fixative agents. This study aimed to analyze the quality of histological preparations of mouse kidney tissue fixed using green betel leaf extract at various concentrations and to compare them with 10% NBF. This experimental study employed extract concentrations of 70%, 85%, and 100%. Samples consisted of healthy male mouse kidney tissues processed through histotechnical procedures and stained using Hematoxylin-Eosin. The quality of the preparations was assessed based on nuclear clarity, cytoplasmic detail, and staining uniformity, followed by statistical analysis using Kruskal Wallis and Mann–Whitney tests. The Mann–Whitney post hoc test showed that the 100% concentration was not significantly different from 10% NBF (p = 0.056; p > 0.05), but was significantly different from 70% (p = 0.002) and 85% (p = 0.007) concentrations (p < 0.05). It can be concluded that green betel leaf extract has potential as a natural alternative fixative, particularly at 100% concentration, as it produces histological quality comparable to the control.