Label-free detection of fibrotic focus in breast tumor microenvironment via multiphoton imaging

Fibrotic focus, a critical pathological feature within tumor microenvironment, is closely correlated with tumor invasiveness and malignancy. Accurately identifying fibrotic focus is therefore essential for early diagnosis, treatment strategy and prognostic assessment of breast cancer. Multiphoton microscopy (MPM) combining two-photon excitation fluorescence (TPEF) and second-harmonic generation (SHG) enables real-time, label-free visualization of pathological changes in tissue slices with subcellular resolution—eliminating the need for staining. In this study, we utilized MPM for label-free detection of fibrotic focus in breast cancer, followed by image data processing to extract collagen features. Our results indicate that MPM is able to rapidly and accurately identify this pathological feature within the tumor microenvironment without the use of exogenous contrast agents. Additionally, quantitative analysis of collagen characteristics provides valuable insights into varying degrees of fibrosis. With its rapid imaging capability and minimal tissue damage, MPM holds significant promise as a diagnostic tool for label-free monitoring of pathological features across various diseases.