Label‐Free Detection of Breast Phyllodes Tumors Based on Multiphoton Microscopy

Phyllodes tumors (PTs) are rare breast stroma neoplasms, and their accurate identification at various stages is essential for personalized patient treatment. In this study, multiphoton microscopy (MPM) with two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) imaging was used for label-free detection and differentiation of PTs and normal breast tissue. An automated image processing strategy was developed to quantify changes in collagen fiber morphology within the stroma and boundary of PTs, establishing optical diagnostic characteristics of PTs using MPM. The results demonstrated that MPM could be used for the detection of different stages of PTs, and the morphological alterations in collagen fibers could serve as critical indicators of PT malignancy, offering new insights for the diagnosis and grading of benign, borderline, and malignant PTs. It lays the groundwork for the future application of compact MPM for the rapid detection and diagnosis of PTs.