<![CDATA[Xylene is a commonly used clearing agent in histological slide preparation; however, its toxic properties necessitate the development of safer alternative materials. One potential alternative is eucalyptus oil (Melaleuca cajuputi). This study aimed to analyze the potential of eucalyptus oil as an alternative clearing agent to replace xylene based on the quality of mouse liver (Mus musculus) histological slides stained with Hematoxylin-Eosin (HE). This experimental study was conducted from October 2025 to May 2026 at PUSVETMA Surabaya, the Faculty of Veterinary Medicine, Universitas Airlangga, and the Cytology and Histotechnology Laboratory of Poltekkes Kemenkes Surabaya. A total of 32 mouse liver histological slides were divided into two groups, consisting of 16 slides cleared using xylene and 16 slides cleared using eucalyptus oil. Slide quality was assessed based on nuclear characteristics, cytoplasm, intercellular boundaries, and color uniformity using a scoring system ranging from 1 to 3. Data were analyzed using the Shapiro-Wilk test, followed by the Kruskal-Wallis and Mann-Whitney tests. The results showed that the mean quality score was 11.13 in the xylene group and 9.44 in the eucalyptus oil group, with a mean difference of 1.69 points. The Mann-Whitney test indicated a significant difference in slide quality between the two groups (p < 0.001). Eucalyptus oil was able to produce histological slides categorized as good, although its quality remained lower than that of xylene as the gold standard. Therefore, eucalyptus oil has the potential to be used as a safer and more environmentally friendly alternative clearing agent.]]>
