Comparison of toluidine blue 1% staining patterns in cytopathologically confirmed ocular surface squamous neoplasias and in non-neoplastic lesions
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Updated: Jan 25, 2026

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Published on: August 9, 2024
Roshanak Aliakbar Navahi1, Seyed Bagher Hosseini2, Mozhgan Rezaei Kanavi2
1Eye Research Center, Rassoul Akram Hospital, Iran University of Medical Sciences, Tehran, Iran.
Purpose:
To evaluate the role of toluidine blue (TB) staining patterns in diagnosis of ocular surface squamous neoplasia (OSSN) in comparison to that of impression cytology.
Methods:
TB 1% dye was applied to different ocular surface lesions, followed by impression cytology (IC). Dye distribution, intensity, and pattern of uptake by the lesion were scored and total score ≥5 was considered "positive TB staining". The TB results were then compared with those using IC to determine the presence of cellular atypia.
Results:
The study enrolled 88 eyes of 82 patients. IC demonstrated cellular atypia in 50 (56.8%) cases. Forty-three of 45 "TB-positive" eyes (95.51%) had cellular atypia on IC (p < 0.001). Sensitivity and specificity of TB in identifying OSSN were 86% and 94.74%, respectively, with positive and negative predictive values of 95.56% and 83.72%. TB staining intensity of dark blue and/or mixed types and stippled pattern of TB staining were strongly correlated with dysplastic changes in IC (P ˂ 0.001). TB staining distribution whether in form of diffuse, patchy or scattered eyes with atypia did not significantly differ from those without atypia in IC (P = 0.172).
Conclusion:
The sensitivity and specificity of TB vital dye in diagnosing OSSN can be increased by focusing on color intensity and a stippled pattern.
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