Abstract
Cervical intraepithelial neoplasia (CIN) represents a spectrum of premalignant lesions requiring accurate early detection to prevent progression to invasive cervical cancer. Colposcopy with visual inspection using acetic acid (VIA) is the gold standard for CIN assessment but suffers from substantial interobserver variability, limiting diagnostic consistency. We evaluated hyperspectral imaging (HSI) as an objective, non-invasive method for characterizing CIN-related tissue changes. This prospective proof-of-principle clinical study enrolled women with histologically confirmed CIN3 indicated for large-loop excision of the transformation zone (LLETZ). Standardized colposcopic images following VIA were obtained and annotated independently by five certified colposcopists according to IFCPC Rio 2011 classification. These annotations served as pathological tissue region references and were quantitatively assessed using intersection over union metrics to evaluate interobserver agreement. HSI was performed immediately prior to LLETZ using the TIVITA Tissue System, capturing spectral reflectance data across 500-995 nm in 100 wavelength bands. Spatial correspondence between colposcopic and hyperspectral images was achieved through homography transformation based on landmark alignment, allowing expert annotations to be projected into the HSI domain. Reflectance spectra from annotated areas were averaged to calculate four proprietary HSI-derived tissue indices, which revealed significantly higher values in CIN-affected regions compared to healthy tissue (p <0.01, Wilcoxon signed-rank test), suggesting increased vascularization and water content. Our findings highlight conventional colposcopy limitations due to examiner subjectivity and support HSI's potential to provide reproducible, quantitative biomarkers for CIN. HSI integration into clinical workflows may enhance cervical cancer screening objectivity and enable reliable diagnostics in resource-limited settings. Clinical and Translational Impact Statement- Hyperspectral imaging enables objective detection of cervical intraepithelial neoplasia and could improve diagnostic accuracy while reducing unnecessary biopsies.