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Cytological diagnosis of follicular thyroid carcinoma (FTC) is one of majorchallenges in the field of endocrine oncology due to absence of evidentmorphological indicators. Morphological abnormalities in the nucleus aretypically key indicators in cancer cytopathology and are attributed to a rangeof biochemical alterations in nuclear components. Consequently, Ramanspectroscopy has been widely used to detect cancer in various cytologicalsamples, often identifying biochemical changes prior to observablemorphological alterations. However, in the case of FTC, cytoplasmic featuressuch as carotenoids, cytochromes, and lipid droplets have shown greaterdiagnostic relevance compared to nuclear features. This study leveragessingle-cell Raman imaging to explore the spatial origin of diagnostic signalsin FTC and normal thyroid (NT) cells, assessing the contributions of thenucleus and cytoplasm independently. Our results demonstrate that Raman spectrafrom the cytoplasmic region can distinguish between FTC and NT cells with anaccuracy of 84% under co-culture conditions, maintaining robustness acrossmultiple devices. In contrast, classification based on nuclear spectra achievedonly 53% accuracy, suggesting that biochemical alterations in the cytoplasmplay a more significant role in FTC detection than those in the nucleus. Ourwork elevates the promise of Raman-based cytopathology by providingcomplementary organelle-dependent information to traditional diagnostic methodsand demonstrating transferability across different devices.

Beyond the Nucleus: Cytoplasmic Dominance in Follicular Thyroid Carcinoma Detection Using Single-Cell Raman Imaging Across Multiple Devices