COMPARATIVE EVALUATION OF KUMKUM VERSUS EOSIN AS A COUNTERSTAIN IN HISTOPATHOLOGICAL SECTIONS OF ORAL LESIONS

Background: Counterstains like eosin are essential in hematoxylin and eosin (H&E) staining for highlighting cytoplasmic and extracellular structures in histopathological analysis. However, synthetic dyes raise concerns over toxicity and cost, prompting exploration of natural alternatives such as Kumkum (Crocosatin from saffron). This study compares Kumkum and eosin as counterstains in oral lesion sections. Objective: To evaluate staining efficacy, color contrast, tissue preservation, and diagnostic utility of Kumkum versus eosin. Materials and Methods: Fifty formalin-fixed, paraffin-embedded sections from oral lesions (e.g., squamous cell carcinoma, leukoplakia, lichen planus) were stained with hematoxylin followed by either 1% aqueous eosin Y or 0.5% Kumkum extract. Sections were assessed blindly by three pathologists for nuclear-cytoplasmic contrast, color stability (under light microscopy at 48 hours), staining intensity (semi-quantitative score 0-4), and diagnostic agreement using kappa statistics. Statistical analysis used paired t-tests and chi-square tests (p<0.05 significant). Results: Kumkum provided comparable nuclear-cytoplasmic contrast (mean score 3.8 ± 0.4 vs. 3.9 ± 0.3 for eosin; p=0.42) with vibrant red-orange hues and superior cytoplasmic eosinophilia in keratinized areas. Color stability was equivalent (fading <5% at 48 hours), and interobserver agreement was high (kappa=0.87 for Kumkum vs. 0.89 for eosin). Kumkum was cost-effective) and non-toxic. Conclusion: Kumkum serves as an effective, economical, natural counterstain alternative to eosin in oral histopathology, with no compromise in diagnostic quality. It holds promise for resource-limited settings.

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Publication Details

Journal
Zenodo (CERN European Organization for Nuclear Research)
Published
2026-03-16
DOI
https://doi.org/10.5281/zenodo.19081851
Primary Topic
Oral Health Pathology and Treatment
Type
article
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article

COMPARATIVE EVALUATION OF KUMKUM VERSUS EOSIN AS A COUNTERSTAIN IN HISTOPATHOLOGICAL SECTIONS OF ORAL LESIONS

Harish Srinivasan1*, Barath Nithish Kumar2, Priyanka Saravanan3, Yogeshwaran S.4, Dhanush S.5, Arnold Allan A.6, Senthilkumar Krishnan7, Gnansekaran8, Rajalingam D.9, Kannabirran Vaikundam10
Zenodo (CERN European Organization for Nuclear Research)
Oral Health Pathology and Treatment
article

COMPARATIVE EVALUATION OF KUMKUM VERSUS EOSIN AS A COUNTERSTAIN IN HISTOPATHOLOGICAL SECTIONS OF ORAL LESIONS

Harish Srinivasan1*, Barath Nithish Kumar2, Priyanka Saravanan3, Yogeshwaran S.4, Dhanush S.5, Arnold Allan A.6, Senthilkumar Krishnan7, Gnansekaran8, Rajalingam D.9, Kannabirran Vaikundam10
article en

Abstract

Background: Counterstains like eosin are essential in hematoxylin and eosin (H&E) staining for highlighting cytoplasmic and extracellular structures in histopathological analysis. However, synthetic dyes raise concerns over toxicity and cost, prompting exploration of natural alternatives such as Kumkum (Crocosatin from saffron). This study compares Kumkum and eosin as counterstains in oral lesion sections. Objective: To evaluate staining efficacy, color contrast, tissue preservation, and diagnostic utility of Kumkum versus eosin. Materials and Methods: Fifty formalin-fixed, paraffin-embedded sections from oral lesions (e.g., squamous cell carcinoma, leukoplakia, lichen planus) were stained with hematoxylin followed by either 1% aqueous eosin Y or 0.5% Kumkum extract. Sections were assessed blindly by three pathologists for nuclear-cytoplasmic contrast, color stability (under light microscopy at 48 hours), staining intensity (semi-quantitative score 0-4), and diagnostic agreement using kappa statistics. Statistical analysis used paired t-tests and chi-square tests (p<0.05 significant). Results: Kumkum provided comparable nuclear-cytoplasmic contrast (mean score 3.8 ± 0.4 vs. 3.9 ± 0.3 for eosin; p=0.42) with vibrant red-orange hues and superior cytoplasmic eosinophilia in keratinized areas. Color stability was equivalent (fading <5% at 48 hours), and interobserver agreement was high (kappa=0.87 for Kumkum vs. 0.89 for eosin). Kumkum was cost-effective) and non-toxic. Conclusion: Kumkum serves as an effective, economical, natural counterstain alternative to eosin in oral histopathology, with no compromise in diagnostic quality. It holds promise for resource-limited settings.

Zenodo (CERN European Organization for Nuclear Research)
Openalex Percentile: Top 17%
Oral Health Pathology and Treatment
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