Alternatives to eosin in histopathological assessments: A comparison of organic stains

Hematoxylin and eosin (H&E) remains the cornerstone of diagnostic pathology, but the potential harmful effects, high cost, and environmental pollution have necessitated the exploration of eco-friendly and economical alternatives.

This study evaluated and compared the overall staining efficacy, durability, and economic feasibility of H&E, hematoxylin and Allium cepa (H&AC), and hematoxylin and Bixa Orellana (H&BO).

The comparative study was conducted in the Department of Oral Pathology and Microbiology, Faculty of Dental Science, Dharmsinh Desai University, Nadiad, Gujarat, over 18 months.

A total of 225 slides, including epithelium, collagen fibers, adipose tissue, salivary gland, and bone, were stained with H&E, H&AC, and H&BO. The overall efficacy evaluation was performed on the basis of staining intensity, clarity, and uniformity.

Staining parameters were graded and statistically analyzed using Chi-square and post hoc Dunn and Bonferroni tests (P < 0.05).

H&E demonstrated superior staining quality across all parameters, with 98.67% excellent intensity, 100% clarity, and 97.33% uniformity (P < 0.001). H&BO emerged as a viable alternative with consistent performance and high efficacy, particularly in collagen and salivary gland tissues. H&AC, while the most cost-effective and eco-friendly, showed variability and reduced staining precision.

While H&E remains the benchmark in diagnostic histopathology, H&BO presents a promising eco-conscious alternative. H&AC, despite its affordability, requires refinement for consistent diagnostic use. This study highlights the potential for balancing diagnostic excellence with environmental responsibility in routine staining protocols.