Kualitas Preparat Histologis Ginjal, Hepar, dan Pankreas Tikus Sprague Dawley Setelah Fiksasi Menggunakan Formalin 10% Selama Dua Minggu
DOI:
https://doi.org/10.30742/kp751w77Keywords:
Fixation, Histotechnical, Kidney, Liver, PancreasAbstract
Background: Histotechnology is a series of procedures used to prepare histological specimens for microscopic analysis. One of the critical steps in this process is fixation, which serves to preserve tissue structure and prevent changes in shape or size. However, excessive fixation duration may lead to over-hardening of tissues, resulting in reduced quality of histological observations. Objectives: To evaluate the feasibility of two week formalin 10% fixation period on histological quality of kidney, liver, and pancreas tissues. Methods: After a 14-day adaptation period, male Sprague Dawley rats were anesthetized, necropsied, and their kidney, liver, and pancreas were fixed in 10% formalin. Histotechnical procedures included graded alcohol dehydration, toluene clearing, paraffin embedding, 6 µm sectioning, and HE staining. Preparations were observed using an Olympus BX41 microscope at magnifications of 4x–40x. Results: Two-week fixation caused structural damage to the kidney, liver, and pancreas. Damage observed included glomeruli and tubules in the kidney, the portal triad structure in the liver, and the Langerhans islet cells in the pancreas. Statistical results (if any) should be included here. Conclusions: Fixation for two weeks did not produce adequate histological preparations, and therefore, it is not recommended as a reference for developing histotechnology SOPs in related laboratories.
References
Adeniran, B. V., Bjarkadottir, B. D., Appeltant, R., Lane, S., & Williams, S. A. (2021). Improved preservation of ovarian tissue morphology that is compatible with antigen detection using a fixative mixture of formalin and acetic acid. Human reproduction (Oxford, England), 36(7), 1871–1890. https://doi.org/10.1093/humrep/deab075
Alturkistani, H. A., Tashkandi, F. M., & Mohammedsaleh, Z. M. (2015). Histological Stains: A Literature Review and Case Study. Global Journal of Health Science, 8(3), 72–79. https://doi.org/10.5539/gjhs.v8n3p72
Anil. Rajendran. (2012). Routine Histotechniques, Staining and Notes on Immunohistochemistry. Shafer’s Textbook of Oral Pathology, January 2012, 944.
Ariany, D., Rahman, A., Rachmawati, N. M., Hermawati, L., & Irawati, N. B. U. (2025). Studi Histopatologi Pengaruh Pemberian Dmba Secara Topikal Pada Organ Hati Mencit C3H/HEJ. Medika Tadulako, 10(1), 66-71. https://doi.org/10.22487/mtj.v10i1.1892
Baskin, D. G. (2014). Fixation and Tissue Processing in Immunohistochemistry. In Pathobiology of Human Disease: A Dynamic Encyclopedia of Disease Mechanisms. Elsevier Inc. https://doi.org/10.1016/B978-0-12-386456-7.07402-5
Bauer, D. R., & Chafin, D. R. (2023). Assessing Tissue Fixation Time and Quality with Label-free Mid Infrared Spectroscopy and Machine Learning. Biopreservation and Biobanking, 21(2), 208–216. https://doi.org/10.1089/bio.2022.0108
Bilgin, C., Dai, D., Johnson, C., Mereuta, O. M., Kallmes, D. F., Brinjikji, W., & Kadirvel, R. (2023). Quality assessment of histopathological stainings on prolonged formalin fixed thrombus tissues retrieved by mechanical thrombectomy. Frontiers in neurology, 14, 1223947. https://doi.org/10.3389/fneur.2023.1223947
Bussolati, G. (2022). Fixation in histopathology: The mandate to renew. Pathologica, 114(4), 275–277. https://doi.org/10.32074/1591-951X-782
Chung, J. Y., Song, J. S., Ylaya, K., Sears, J. D., Choi, L., Cho, H., Rosenberg, A. Z., & Hewitt, S. M. (2018). Histomorphological and Molecular Assessments of the Fixation Times Comparing Formalin and Ethanol-Based Fixatives. Journal of Histochemistry and Cytochemistry, 66(2), 121–135. https://doi.org/10.1369/0022155417741467
Dağdeviren, T., Yolcu, H. K., & Ünver, B. (2024). Histological Fixation Process and Fixatives. Turkish Journal of Agriculture - Food Science and Technology, 12(8), 1482–1486. https://doi.org/10.24925/turjaf.v12i8.1482-1486.6808
Likhithaswamy, H. R., Madhushankari, G. S., Selvamani, M., Mohan Kumar, K. P., Kokila, G., & Mahalakshmi, S. (2022). Assessing the quality of long-term stored tissues in formalin and in paraffin-embedded blocks for histopathological analysis. Journal of Microscopy and Ultrastructure, 10(1), 23–29. https://doi.org/10.4103/JMAU.JMAU_53_20
Lim, S. D., Huang, Q., & Seibel, E. J. (2019). Evaluation of Formalin Fixation for Tissue Biopsies Using Shear Wave Laser Speckle Imaging System. IEEE Journal of Translational Engineering in Health and Medicine, 7(March), 1–10. https://doi.org/10.1109/JTEHM.2019.2909914
Meecham, A., Miranda, E., Morris, H. T., Hair, J., Oien, K. A., Gerrard, G., Guppy, N., Mooney, D., Shaw, E. C., Ashton-Key, M., Lees, R., Flanagan, A., & Rodriguez-Justo, M. (2021). Alternative tissue fixation for combined histopathological and molecular analysis in a clinically representative setting. Histochemistry and Cell Biology, 156(6), 595–607. https://doi.org/10.1007/s00418-021-02029-1
Morrison, L. E., Larrinaga, T. M., Kelly, B. D., Lefever, M. R., Beck, R. C., & Bauer, D. R. (2025). Concurrent Viewing of H&E and Multiplex Immunohistochemistry in Clinical Specimens. Diagnostics, 15(2). https://doi.org/10.3390/diagnostics15020164
Olenovych, O., Boychuk, T., Davydenko, I. S., & Davydenko, O. (2024). Histomorphological Peculiarities of the Pancreatic Parenchyma in Rats With Alloxan-Induced Diabetes of Different Duration. Neonatology, Surgery and Perinatal Medicine, 14(2(52)), 100–107. https://doi.org/10.24061/2413-4260.XIV.2.52.2024.15
Rahman, M. A., Sultana, N., Ayman, U., Bhakta, S., Afrose, M., Afrin, M., & Haque, Z. (2022). Alcoholic fixation over formalin fixation: A new, safer option for morphologic and molecular analysis of tissues. Saudi journal of biological sciences, 29(1), 175–182. https://doi.org/10.1016/j.sjbs.2021.08.075
Sanderson, T., Wild, G., Cull, A. M., Marston, J., & Zardin, G. (2018). Immunohistochemical and immunofluorescent techniques. Bancroft’s Theory and Practice of Histological Techniques, Eighth Edition, 5, 337–394. https://doi.org/10.1016/B978-0-7020-6864-5.00019-0
Shami, G. J., Chen, Z., Cheng, D., Wisse, E., & Braet, F. (2025). On the long-term storage of tissue for fluorescence and electron microscopy: lessons learned from rat liver samples. Histochemistry and Cell Biology, 163(1). https://doi.org/10.1007/s00418-024-02334-5
Shetty, J. K., Babu, H. F., & Hosapatna Laxminarayana, K. P. (2020). Histomorphological Assessment of Formalin versus Nonformalin Fixatives in Diagnostic Surgical Pathology. Journal of Laboratory Physicians, 12(04), 271–275. https://doi.org/10.1055/s-0040-1722546
Silva, J., Castelhano, R., Silva, F., Santos, J. P., Félix, A., Cruz, J., Machado, J., & Pessanha, S. (2024). The impact of formalin fixation in the elemental content of tissues: Parametrization up to 48 h. Microchemical Journal, 200(February). https://doi.org/10.1016/j.microc.2024.110281
Tran, H., Jan, N. J., Hu, D., Voorhees, A., Schuman, J. S., Smith, M. A., Wollstein, G., & Sigal, I. A. (2017). Formalin Fixation and Cryosectioning Cause Only Minimal Changes in Shape or Size of Ocular Tissues. Scientific reports, 7(1), 12065. https://doi.org/10.1038/s41598-017-12006-1
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Copyright (c) 2025 Muhammad Azharan Alwi, Nurlaely Mida Rachmawati, Devy Ariany, Luluk Hermawati, Hilizza Awalina Zulfa
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