The Impact Of Histological Tissue Processing On Diagnostic Accuracy In Histopathology: A Narrative Review

Rhemadhyaksa Tegar Arrondiwa; Khairun Nisa; Pratiwi Soesilawati; Zahiyah Wulan Supriyono; Valerian Rayhan Wijaya

doi:10.37841/jidam.2026.v13.i3.006

Review

Article Type

5

Authors

26

References

📝Abstract

📥 Received: 8 August 2026 ✅ Accepted: 25 August 2026 📤 Published: 27 September 2026

Background/Aim: Histopathological diagnosis depends on high-quality tissue processing to preserve tissue morphology and ensure diagnostic accuracy. Inadequate processing may induce artifacts and compromise diagnostic interpretation. This narrative review aimed to summarize current evidence on the impact of histological tissue processing on tissue quality, artifact formation, and diagnostic accuracy.

Methods: A narrative review was conducted using a structured systematic literature search guided by the PRISMA 2020 statement. The review question was formulated using the PEO framework. English-language studies published between 2021 and 2026 were retrieved from PubMed, Scopus, and ScienceDirect using predefined inclusion and exclusion criteria, and the eligible studies were synthesized narratively.

Results: FFPE remains the gold standard for preserving tissue morphology and structural integrity. However, processing-related artifacts and formalin-induced molecular alterations may reduce diagnostic accuracy. Emerging approaches, including alternative processing reagents, microwave-assisted and automated processing, and artificial intelligence-assisted quality control, show promise for improving specimen quality and diagnostic reliability.

Conclusion: Optimizing and standardizing histological tissue processing is essential to improve diagnostic accuracy and support advances in molecular pathology, digital pathology, and precision medicine.

Keywords: Histology Paraffin Embedding Formaldehyde Artifacts Diagnostic Accuracy Digital Pathology

📋How to Cite

Arrondiwa RT, Nisa K, Soesilawati P, Supriyono ZW, Wijaya VR. The impact of histological tissue processing on diagnostic accuracy in histopathology: a narrative review. J Indian Dent Assoc Madras. 2026;13(3):31-45. DOI: 10.37841/jidam.2026.v13.i3.006

Arrondiwa, R. T., Nisa, K., Soesilawati, P., Supriyono, Z. W., & Wijaya, V. R. (2026). The impact of histological tissue processing on diagnostic accuracy in histopathology: A narrative review. Journal of Indian Dental Association Madras, 13(3), 31–45. https://doi.org/10.37841/jidam.2026.v13.i3.006

Arrondiwa, Rhemadhyaksa Tegar, et al. "The Impact Of Histological Tissue Processing On Diagnostic Accuracy In Histopathology: A Narrative Review." Journal of Indian Dental Association Madras, vol. 13, no. 3, 2026, pp. 31–45. DOI: 10.37841/jidam.2026.v13.i3.006.

Arrondiwa RT, Nisa K, Soesilawati P, Supriyono ZW, Wijaya VR. The impact of histological tissue processing on diagnostic accuracy in histopathology: a narrative review. J Indian Dent Assoc Madras. 2026;13(3):31-45. doi:10.37841/jidam.2026.v13.i3.006

Arrondiwa, Rhemadhyaksa Tegar, Khairun Nisa, Pratiwi Soesilawati, Zahiyah Wulan Supriyono, and Valerian Rayhan Wijaya. "The Impact Of Histological Tissue Processing On Diagnostic Accuracy In Histopathology: A Narrative Review." Journal of Indian Dental Association Madras 13, no. 3 (2026): 31–45. https://doi.org/10.37841/jidam.2026.v13.i3.006.

📚References

1
Bright OA, Samuel DN, Adisa MA, Dorcas OO, Perez Q, Melody AA, Michael AK, Albert OS, Senu E. Clearing Properties Between Coconut Oil and Xylene in Histological Tissue Processing. J Histochem Cytochem. 2024;72(4):233-243. doi: 10.1369/00221554241241369.
2
Gundlapalle P, Nagappan N, Ramesh P, Ziauddhin S, Karthick BP, Paleti VS, Kumar BV. Comparison of Oral Mucosal Biopsies Done Using Scalpel and Diode Lasers: An In Vivo Study. J Pharm Bioallied Sci. 2022;14(Suppl 1):S947-S954. doi: 10.4103/jpbs.jpbs_861_21.
3
Falahkheirkhah K, Guo T, Hwang M, Tamboli P, Wood CG, Karam JA, Sircar K, Bhargava R. A generative adversarial approach to facilitate archival-quality histopathologic diagnoses from frozen tissue sections. Lab Invest. 2022;102(5):554-559. doi: 10.1038/s41374-021-00718-y.
4
Cavalcanti E, et al. A new reliable method for tissue preservation. Pathol Res Pract. 2022;234:153910. doi: 10.1016/j.prp.2022.153910.
5
Aziz SJ, Zeman-Pocrnich CE. Tissue Processing. Methods Mol Biol. 2022;2422:47-63. doi: 10.1007/978-1-0716-1948-3_4.
6
Haghbin N, Oveisi B, Banitaba AP. Automated variable power cold microwave tissue processing: A novel universal tissue processing protocol without using formaldehyde and xylene. Acta Histochem. 2022;124(4):151880. doi: 10.1016/j.acthis.2022.151880.
7
Kanwal N, López-Pérez M, Kiraz U, Zuiverloon TCM, Molina R, Engan K. Are you sure it's an artifact? Artifact detection and uncertainty quantification in histological images. Comput Med Imaging Graph. 2024;112:102321. doi: 10.1016/j.compmedimag.2023.102321.
8
Flint N, Friedman P, Frigerio A, Tinklepaugh A. Incomplete Staining Artifact: A Confounding Frozen Section Pathology Artifact Encountered During Mohs Micrographic Surgery. J Drugs Dermatol. 2022;21(5):542-544. doi: 10.36849/JDD.6722.
9
Weng Z, Seper A, Pryalukhin A, et al. GrandQC: A comprehensive solution to quality control problem in digital pathology. Nat Commun. 2024;15:10582. doi: 10.1038/s41467-024-54769-y.
10
Wang C, Qu K, Li S, Yu Y, He J, Zhang C, Shen Y. ArtiDiffuser: A unified framework for artifact restoration and synthesis for histology images via counterfactual diffusion model. Med Image Anal. 2025;102:103567. doi: 10.1016/j.media.2025.103567.
11
Olave MC, Manrai P, Nalbantoglu I, Jain D, Barbieri AL. Push pin induced artifact: A mimic of iron pill injury in gastrointestinal pathology. Hum Pathol. 2026;173:106119. doi: 10.1016/j.humpath.2026.106119.
12
Tay N, Alshammari A, Kaur S, Pettit A, Mu E, Reid A, Winkler I, Vetter I, Starobova H. A comprehensive protocol for simplified mouse DRG fixation, processing and immunohistochemistry: Overcoming common challenges. J Neurosci Methods. 2025;418:110434. doi: 10.1016/j.jneumeth.2025.110434.
13
Rahman MA, Sultana N, Ayman U, Bhakta S, Afrose M, Afrin M, Haque Z. Alcoholic fixation over formalin fixation: A new, safer option for morphologic and molecular analysis of tissues. Saudi J Biol Sci. 2022;29(1):175-182. doi: 10.1016/j.sjbs.2021.08.075.
14
De Martino E, Medeot C, D'Amico L, Stanta G, Bonin S. Impact of standardization in tissue processing: the performance of different fixatives. N Biotechnol. 2022;71:30-36. doi: 10.1016/j.nbt.2022.07.001.
15
Villanacci V, Del Sordo R, Casella G, Becheanu G, Oberti A, Belfekih B, Bassotti G, Ravelli A. The correct methodological approach to the diagnosis of celiac disease: the point of view of the pathologist. Gastroenterol Hepatol Bed Bench. 2023;16(2). doi: 10.22037/ghfbb.v16i2.2704.
16
Borges MA, Curcio BR, Gastal GDA, Gheno L, Varela Junior AS, Corcini CD, Nogueira CEW, Aguiar FLN, Gastal EL. Ethanol, Carnoy, and paraformaldehyde as fixative solutions for histological evaluation of preantral follicles in equine ovarian tissue. Reprod Biol. 2023;23(4):100814. doi: 10.1016/j.repbio.2023.100814.
17
Hacihasanoglu E, et al. PD-L1 Assessment in Needle Core Biopsies of Non-Small Cell Lung Cancer: Interpathologist Agreement and Potential Associated Histopathological Features. Turk J Pathol. 2024. doi: 10.5146/tjpath.2023.01609.
18
Hamed A, Hoskuldsson T, Aldaraiseh B, Alutaibi M, Karzoun MZ, Jamshed A, Paz M, Hamed N, Cartwright D, Turner KO, Khalifa MA, Amin K, Park BU. Thermal resection-margin artifact mimicking goblet cell adenocarcinoma in appendectomy specimens: clinicopathologic correlation with appendiceal stump technique. Hum Pathol. 2026;175:106157. doi: 10.1016/j.humpath.2026.106157.
19
Winter L, Schmitt VH, Barsch F, Schwarz D, Cotarelo CL, Brochhausen C. Comparative Analysis of Histomorphological Quality and Quantitative Cell Assessment in Formalin-Fixed Paraffin-Embedded and Fresh Frozen Porcine Skin Biopsies. Biomedicines. 2026;14(2):390. doi: 10.3390/biomedicines14020390.
20
Ghebeh H, Mansour FA, Colak D, Alfuraydi AA, Al-Thubiti AA, Monies D, Al-Alwan M, Al-Tweigeri T, Tulbah A. Higher PD-L1 Immunohistochemical Detection Signal in Frozen Compared to Matched Paraffin-Embedded Formalin-Fixed Tissues. Antibodies. 2021;10(3):24. doi: 10.3390/antib10030024.
21
Basting RT, Napimoga MH, Lima JM, Freitas NS, Clemente-Napimoga JT. Fast and accurate protocol for histology and immunohistochemistry reactions in temporomandibular joint of rats. Arch Oral Biol. 2021;126:105115. doi: 10.1016/j.archoralbio.2021.105115.
22
Yadav RP, Xing P, Chen X. scFFPE-ATAC enables high-throughput single cell chromatin accessibility profiling in formalin-fixed paraffin-embedded samples. Nat Commun. 2025;16:66170. doi: 10.1038/s41467-025-66170-4.
23
Oliveira DVNP, Hentze J, et al. DNA Methylation in Ovarian Tumors — A Comparison Between Fresh and FFPE. Reprod Sci. 2021;28:3212-3218. doi: 10.1007/s43032-021-00589-0.
24
Ong FN, Aye SN, Krishnappa P, Ravindran R. A comparative study of microwave oven-assisted tissue processing and conventional method of tissue processing on turnaround laboratory time and morphological quality of tissue sections. Med J Malaysia. 2023. PMID: 36988531.
25
Thangamani SS, Ravindran S, Thiagarajan MK, Mayilvakanam SK, Ravi A, Annamalai S. Rethinking tissue preservation: A review of non-toxic and environmentally sustainable fixatives. Ann Diagn Pathol. 2025;78:152500. doi: 10.1016/j.anndiagpath.2025.152500.
26
Wiedenmann M, Barch M, Chang PS, Giltnane J, Risom T, Zijlstra A. An Immunofluorescence-Guided Segmentation Model in Hematoxylin and Eosin Images Is Enabled by Tissue Artifact Correction Using a Cycle-Consistent Generative Adversarial Network. Mod Pathol. 2024;37(6):100591. doi: 10.1016/j.modpat.2024.100591.

[1] 1
Bright OA, Samuel DN, Adisa MA, Dorcas OO, Perez Q, Melody AA, Michael AK, Albert OS, Senu E. Clearing Properties Between Coconut Oil and Xylene in Histological Tissue Processing. J Histochem Cytochem. 2024;72(4):233-243. doi: 10.1369/00221554241241369.

[2] 2
Gundlapalle P, Nagappan N, Ramesh P, Ziauddhin S, Karthick BP, Paleti VS, Kumar BV. Comparison of Oral Mucosal Biopsies Done Using Scalpel and Diode Lasers: An In Vivo Study. J Pharm Bioallied Sci. 2022;14(Suppl 1):S947-S954. doi: 10.4103/jpbs.jpbs_861_21.

[3] 3
Falahkheirkhah K, Guo T, Hwang M, Tamboli P, Wood CG, Karam JA, Sircar K, Bhargava R. A generative adversarial approach to facilitate archival-quality histopathologic diagnoses from frozen tissue sections. Lab Invest. 2022;102(5):554-559. doi: 10.1038/s41374-021-00718-y.

[4] 4
Cavalcanti E, et al. A new reliable method for tissue preservation. Pathol Res Pract. 2022;234:153910. doi: 10.1016/j.prp.2022.153910.

[5] 5
Aziz SJ, Zeman-Pocrnich CE. Tissue Processing. Methods Mol Biol. 2022;2422:47-63. doi: 10.1007/978-1-0716-1948-3_4.

[6] 6
Haghbin N, Oveisi B, Banitaba AP. Automated variable power cold microwave tissue processing: A novel universal tissue processing protocol without using formaldehyde and xylene. Acta Histochem. 2022;124(4):151880. doi: 10.1016/j.acthis.2022.151880.

[7] 7
Kanwal N, López-Pérez M, Kiraz U, Zuiverloon TCM, Molina R, Engan K. Are you sure it's an artifact? Artifact detection and uncertainty quantification in histological images. Comput Med Imaging Graph. 2024;112:102321. doi: 10.1016/j.compmedimag.2023.102321.

[8] 8
Flint N, Friedman P, Frigerio A, Tinklepaugh A. Incomplete Staining Artifact: A Confounding Frozen Section Pathology Artifact Encountered During Mohs Micrographic Surgery. J Drugs Dermatol. 2022;21(5):542-544. doi: 10.36849/JDD.6722.

[9] 9
Weng Z, Seper A, Pryalukhin A, et al. GrandQC: A comprehensive solution to quality control problem in digital pathology. Nat Commun. 2024;15:10582. doi: 10.1038/s41467-024-54769-y.

[10] 10
Wang C, Qu K, Li S, Yu Y, He J, Zhang C, Shen Y. ArtiDiffuser: A unified framework for artifact restoration and synthesis for histology images via counterfactual diffusion model. Med Image Anal. 2025;102:103567. doi: 10.1016/j.media.2025.103567.

[11] 11
Olave MC, Manrai P, Nalbantoglu I, Jain D, Barbieri AL. Push pin induced artifact: A mimic of iron pill injury in gastrointestinal pathology. Hum Pathol. 2026;173:106119. doi: 10.1016/j.humpath.2026.106119.

[12] 12
Tay N, Alshammari A, Kaur S, Pettit A, Mu E, Reid A, Winkler I, Vetter I, Starobova H. A comprehensive protocol for simplified mouse DRG fixation, processing and immunohistochemistry: Overcoming common challenges. J Neurosci Methods. 2025;418:110434. doi: 10.1016/j.jneumeth.2025.110434.

[13] 13
Rahman MA, Sultana N, Ayman U, Bhakta S, Afrose M, Afrin M, Haque Z. Alcoholic fixation over formalin fixation: A new, safer option for morphologic and molecular analysis of tissues. Saudi J Biol Sci. 2022;29(1):175-182. doi: 10.1016/j.sjbs.2021.08.075.

[14] 14
De Martino E, Medeot C, D'Amico L, Stanta G, Bonin S. Impact of standardization in tissue processing: the performance of different fixatives. N Biotechnol. 2022;71:30-36. doi: 10.1016/j.nbt.2022.07.001.

[15] 15
Villanacci V, Del Sordo R, Casella G, Becheanu G, Oberti A, Belfekih B, Bassotti G, Ravelli A. The correct methodological approach to the diagnosis of celiac disease: the point of view of the pathologist. Gastroenterol Hepatol Bed Bench. 2023;16(2). doi: 10.22037/ghfbb.v16i2.2704.

[16] 16
Borges MA, Curcio BR, Gastal GDA, Gheno L, Varela Junior AS, Corcini CD, Nogueira CEW, Aguiar FLN, Gastal EL. Ethanol, Carnoy, and paraformaldehyde as fixative solutions for histological evaluation of preantral follicles in equine ovarian tissue. Reprod Biol. 2023;23(4):100814. doi: 10.1016/j.repbio.2023.100814.

[17] 17
Hacihasanoglu E, et al. PD-L1 Assessment in Needle Core Biopsies of Non-Small Cell Lung Cancer: Interpathologist Agreement and Potential Associated Histopathological Features. Turk J Pathol. 2024. doi: 10.5146/tjpath.2023.01609.

[18] 18
Hamed A, Hoskuldsson T, Aldaraiseh B, Alutaibi M, Karzoun MZ, Jamshed A, Paz M, Hamed N, Cartwright D, Turner KO, Khalifa MA, Amin K, Park BU. Thermal resection-margin artifact mimicking goblet cell adenocarcinoma in appendectomy specimens: clinicopathologic correlation with appendiceal stump technique. Hum Pathol. 2026;175:106157. doi: 10.1016/j.humpath.2026.106157.

[19] 19
Winter L, Schmitt VH, Barsch F, Schwarz D, Cotarelo CL, Brochhausen C. Comparative Analysis of Histomorphological Quality and Quantitative Cell Assessment in Formalin-Fixed Paraffin-Embedded and Fresh Frozen Porcine Skin Biopsies. Biomedicines. 2026;14(2):390. doi: 10.3390/biomedicines14020390.

[20] 20
Ghebeh H, Mansour FA, Colak D, Alfuraydi AA, Al-Thubiti AA, Monies D, Al-Alwan M, Al-Tweigeri T, Tulbah A. Higher PD-L1 Immunohistochemical Detection Signal in Frozen Compared to Matched Paraffin-Embedded Formalin-Fixed Tissues. Antibodies. 2021;10(3):24. doi: 10.3390/antib10030024.

[21] 21
Basting RT, Napimoga MH, Lima JM, Freitas NS, Clemente-Napimoga JT. Fast and accurate protocol for histology and immunohistochemistry reactions in temporomandibular joint of rats. Arch Oral Biol. 2021;126:105115. doi: 10.1016/j.archoralbio.2021.105115.

[22] 22
Yadav RP, Xing P, Chen X. scFFPE-ATAC enables high-throughput single cell chromatin accessibility profiling in formalin-fixed paraffin-embedded samples. Nat Commun. 2025;16:66170. doi: 10.1038/s41467-025-66170-4.

[23] 23
Oliveira DVNP, Hentze J, et al. DNA Methylation in Ovarian Tumors — A Comparison Between Fresh and FFPE. Reprod Sci. 2021;28:3212-3218. doi: 10.1007/s43032-021-00589-0.

[24] 24
Ong FN, Aye SN, Krishnappa P, Ravindran R. A comparative study of microwave oven-assisted tissue processing and conventional method of tissue processing on turnaround laboratory time and morphological quality of tissue sections. Med J Malaysia. 2023. PMID: 36988531.

[25] 25
Thangamani SS, Ravindran S, Thiagarajan MK, Mayilvakanam SK, Ravi A, Annamalai S. Rethinking tissue preservation: A review of non-toxic and environmentally sustainable fixatives. Ann Diagn Pathol. 2025;78:152500. doi: 10.1016/j.anndiagpath.2025.152500.

[26] 26
Wiedenmann M, Barch M, Chang PS, Giltnane J, Risom T, Zijlstra A. An Immunofluorescence-Guided Segmentation Model in Hematoxylin and Eosin Images Is Enabled by Tissue Artifact Correction Using a Cycle-Consistent Generative Adversarial Network. Mod Pathol. 2024;37(6):100591. doi: 10.1016/j.modpat.2024.100591.