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Histology

Histological evaluation of eco-friendly clearing agents as substitutes for xylol in hematoxylin and eosin staining of fibroadenoma mammae tissue

Wimpy Sekolah Tinggi Ilmu Kesehatan Nasional, Sukoharjo, Indonesia, Indonesia Correspondence wimpy@stikesnas.ac.id
,
Endang Widhiyastuti Sekolah Tinggi Ilmu Kesehatan Nasional, Sukoharjo, Indonesia, Indonesia Correspondence dr.endangwidhiyastuti@stikesnas.ac.id
,
Astri Aditya Wardhani Departemen of Patology Anatomi, Rumah Sakit Islam Surakarta, Indonesia, Indonesia Correspondence astriadityawardhani@gmail.com
,
Eva Ocha Pratama Sekolah Tinggi Ilmu Kesehatan Nasional, Sukoharjo, Indonesia, Indonesia Correspondence ramadhanarya570@gmail.com
,
Khoirunisa Saputri Sekolah Tinggi Ilmu Kesehatan Nasional, Sukoharjo, Indonesia, Indonesia Correspondence chnsaaaa62@gmail.com
,
Ardy Prian Nirwana Sekolah Tinggi Ilmu Kesehatan Nasional, Sukoharjo, Indonesia, Indonesia Correspondence ardypriannirwana@stikenas.ac.id
Pages 253-266
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Abstract

Xylol is widely used as a clearing agent in histopathological tissue processing because of its excellent ability to enhance tissue transparency and staining quality. However, its toxicity, volatility, and potential carcinogenicity pose serious occupational and environmental hazards. This cross-sectional analytical experimental study evaluated the histological performance of natural essential oil based clearing agents as sustainable substitutes for xylol in hematoxylin and eosin (H&E) staining of fibroadenoma mammae tissue. Twenty-eight archival samples were processed using α-pinene, 1,8-cineole, or virgin coconut oil (VCO), with xylol serving as the control. Specimens were fixed in 10% neutral buffered formalin, dehydrated in graded alcohol, and evaluated macroscopically and microscopically using an ordinal scoring system encompassing tissue shrinkage, transparency, color uniformity, nuclear and cytoplasmic clarity, staining quality, and integrity. Data were analyzed descriptively and statistically. Results showed that α-pinene and VCO achieved total scores of 32.57 ± 0.30 (p = 0.784) and 32.14 ± 0.26 (p = 0.453), respectively, comparable to xylol (33.00 ± 0.00), while 1,8-cineole performed significantly lower (22.43 ± 0.65, p < 0.05). These findings highlight the novelty of using α-pinene and VCO as effective, non-toxic, inexpensive, and readily available xylol alternatives, reinforcing their relevance for sustainable histopathology, particularly in resource-limited settings.

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How to Cite This

Wimpy, Endang Widhiyastuti, Astri Aditya Wardhani, Eva Ocha Pratama, Khoirunisa Saputri, & Ardy Prian Nirwana. (2025). Histological evaluation of eco-friendly clearing agents as substitutes for xylol in hematoxylin and eosin staining of fibroadenoma mammae tissue. Jurnal Teknologi Laboratorium, 14(2), 253–266. https://doi.org/10.29238/teknolabjournal.v14i2.698

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