Fibroblasts are important players in proliferation, invasion, migration and metastasis of cancer cells. Until now, histological methods such as hematoxylin and eosin (H and E) staining are still a gold standard to assess fibroblasts in traditional histopathology. Multiphoton microscopy (MPM) has become a key technology for biological tissues imaging without staining at the cellular level. In this study, MPM based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) was used to identify the fibroblasts in breast cancer tissue. Normal fibroblasts with spindle shape and abnormal fibroblasts with stellate shape could be clearly obtained by MPM.
Florid type is a kind of morphological variation of lobular carcinoma in situ (LCIS). Florid lobular carcinoma in situ (FLCIS) has the same cytological features as LCIS, often associated with comedo-type necrosis. Unlike classic lobular carcinoma in situ (CLCIS), which is often managed with close observation or chemoprevention, the treatment guidelines recommend that FLCIS be managed in the same way as ductal carcinoma in situ (DCIS). Therefore, it is critical to accurately identify FLCIS for management purposes. Recently, multiphoton microscopy (MPM) has become a powerful imaging tool for label-free detection of biological tissue. In this work, we obtain high-resolution images of unstained normal and diseased breast specimens by MPM. In order to verify the imaging details, we also obtained hematoxylin-eosin (H and E) stained images of corresponding tissues to compare with the MPM images. Our results indicate that MPM can identify FLCIS and CLCIS through histological characteristics, including cell morphology and collagen structure. With the further improvement of MPM, its diagnostic capabilities of real-time and non-invasive may provide a new option for early detection of breast tumor
Perineural invasion (PNI) is the process of neoplastic invasion of nerves. The nerve sheath includes three layers, that is, the epineurium, perineurium and endoneurium from the outside to the inside. Any one of them is invaded by malignant tumor cells, or more than 33% of the circumference of the nerve should be surrounded by tumor cells, then it can be defined as perineural invasion (PNI). Perineural invasion is another way of tumor cell metastasis, which not only indicates the poor prognosis of patients but also indicates the decline of survival rate. In this study, multiphoton microscopy (MPM) which is based on the intrinsic nonlinear optical signals—two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) was applied to identify the perineural invasion in breast cancer patients, including the recognition of normal nerves and tumor cells infiltrating into the nerve. Our aim is to determine the potential feasibility of using this microscopy to identify tumor invasion around the nerves. The results mean that it is feasible to detect perineural invasion with multiphoton microscopy, and to provide a new auxiliary tool for pathologists to quickly and effectively diagnose perineural invasion.
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