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While holotomography enables 3D, label\u2010free imaging of LDs via refractive index, it relies on complex thresholding and lacks biochemical specificity. Here, polarization\u2010sensitive holotomography (PS\u2010HT), which leverages the intrinsic birefringence of LDs for high\u2010contrast, selective identification with a fixed near\u2010zero threshold is presented. Using prostate cell models (healthy PNT2 and cancer PC3), PS\u2010HT is validated against fluorescence microscopy and holotomography, showing that it enables accurate quantification of birefringence, LD volume, dry mass, molecular organization, and spatial distribution. Cancer cells show significantly higher birefringence after glucose treatment, reflecting enhanced lipid accumulation. PS\u2010HT, combined with principal component analysis, achieves near\u2010perfect classification of cancer versus healthy cells, establishing it as a robust, label\u2010free tool for studying lipid metabolism and cancer diagnostics.<\/jats:p>","DOI":"10.1002\/advs.202509420","type":"journal-article","created":{"date-parts":[[2025,8,21]],"date-time":"2025-08-21T08:20:12Z","timestamp":1755764412000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Polarization\u2010Sensitive Holotomography for Multidimensional Label\u2010Free Imaging and Characterization of Lipid Droplets in Cancer Cells"],"prefix":"10.1002","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5360-8261","authenticated-orcid":false,"given":"Hossein","family":"Khadem","sequence":"first","affiliation":[{"name":"Institute of Endotypes in Oncology, Metabolism and Immunology \u201cGaetano Salvatore\u201d IEOMI Second Unit, National Research Council  Via P. 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