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Photon Absorption Remote Sensing Virtual Histopathology: Diagnostic Equivalence to Gold-Standard H&E Staining in Skin Cancer Excisional Biopsies

Photon Absorption Remote Sensing (PARS) enables label-free imaging of subcellular morphology by observing biomolecule specific absorption interactions. Coupled with deep-learning, PARS produces label-free virtual Hematoxylin and Eosin (H&E) stained images in unprocessed tissues. This study evaluates the diagnostic performance of these PARS-derived virtual H&E images in benign and malignant excisional skin biopsies, including Squamous (SCC), Basal (BCC) Cell Carcinoma, and normal skin. Sixteen unstained formalin-fixed paraffin-embedded skin excisions were PARS imaged, virtually H&E stained, then chemically stained and imaged at 40x. Seven fellowship trained dermatopathologists assessed all 32 images in a masked randomized fashion. Concordance analysis indicates 95.5% agreement between primary diagnoses rendered on PARS versus H&E images (Cohen's k=0.93). Inter-rater reliability was near-perfect for both image types (Fleiss' k=0.89 for PARS, k=0.80 for H&E). For subtype classification, agreement was near-perfect 91% (k=0.73) for SCC and was perfect for BCC. When assessing malignancy confinement (e.g., cancer margins), agreement was 92% between PARS and H&E (k=0.718). During assessment dermatopathologists could not reliably distinguish image origin (PARS vs. H&E), and diagnostic confidence was equivalent between the modalities. Inter-rater reliability for PARS virtual H&E was consistent with reported benchmarks for histologic evaluation. These results indicate that PARS virtual histology may be diagnostically equivalent to traditional H&E staining in dermatopathology diagnostics, while enabling assessment directly from unlabeled, or unprocessed slides. In turn, the label-free PARS virtual H&E imaging workflow may preserve tissue for downstream analysis while producing data well-suited for AI integration potentially accelerating and enhancing the accuracy of skin cancer diagnostics.

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