Rosli, Danish Fikri and Mohd Norhadi, Dina Izzati and Ibrahim, Nur Hazreen and Halim, Mohd Dhiyaulhaq and Miskam, Muhammad Faiz and Mohd Radzuan, Hazulin (2026) Enhancing haematoxylin and eosin staining consistency for reproducible histopathology: a study on pH and section thickness. In: 10th International Virtual Medical Research Symposium, 2026, Online.
|
PDF (Abstract)
- Published Version
Download (1MB) | Preview |
Abstract
Introduction: Hematoxylin and eosin (H&E) staining remains fundamental to diagnostic histopathology, medical teaching, and tissue-based research. With increasing reliance on digital pathology and AI-assisted interpretation, maintaining consistent, high-quality staining is critical, as automated systems cannot compensate for poor staining variation. Standardised manual protocols are therefore essential across laboratories. This study investigates how hematoxylin pH and tissue section thickness influence staining quality in loose and compact tissues to support more reproducible H&E workflows. Materials and methods: Duodenum and heart tissue from male Sprague-Dawley rats were fixed in neutral-buffered formalin, processed into paraffin blocks, and sectioned at 3um, 5um and 7um. Sections were stained using hematoxylin adjusted to pH 2.3, 2.6 and 2.9. Nuclear staining intensity was quantified using imageJ. Two-way ANOVA with Tukey's post-hoc test assessed the effects of pH and thickness. Results: Both pH and thickness significantly affected nuclear staining intensity in duodenum (pH: p = 0.008; thickness: p<0.001) and heart (pH: p = 0.0015; thickness: p = 0.001), with no significant interaction. Hematoxylin at pH 2.3 produced weaker nuclear staining, while pH 2.6 and 2.9 improved colour intensity and definition. Thicker sections (7um) consistently showed reduced staining quality. Conclusion: Optimal staining was achieved at pH 2.6 for duodenum and pH 2.9 for heart, with 3-5um sections recommended for reliable outcomes. These findings highlight that consistent manual staining remains fundamental, reinforcing the importance of robust, standardised H&E workflows for reproducible and clinically meaningful histological evaluation.
| Item Type: | Proceeding Paper (Poster) |
|---|---|
| Additional Information: | 6518/127985 |
| Uncontrolled Keywords: | H&E, pH, reproducible staining, section thickness |
| Subjects: | Q Science > Q Science (General) Q Science > QM Human anatomy R Medicine > R Medicine (General) R Medicine > RB Pathology R Medicine > RZ Other systems of medicine |
| Kulliyyahs/Centres/Divisions/Institutes (Can select more than one option. Press CONTROL button): | Kulliyyah of Medicine Kulliyyah of Medicine > Department of Basic Medical |
| Depositing User: | Dr Hazulin Mohd Radzuan |
| Date Deposited: | 18 Mar 2026 16:54 |
| Last Modified: | 18 Mar 2026 16:54 |
| Queue Number: | 2026-03-Q2588 |
| URI: | http://irep.iium.edu.my/id/eprint/127985 |
Actions (login required)
 |
View Item |
Download Statistics
Download Statistics