In recent years, the detection rate of ductal carcinoma in situ (DCIS) has been greatly increased due to the wide application of mammography. DCIS is a non-invasive neoplastic lesion that encompassed a heterogeneous group of lesions characterized by mild to severe atypia in ductal hyperplastic tumor cells. To facilitate the selection of appropriate treatments and to predict the risk of recurrence of DCIS, pathologists divide DCIS into three grades based primarily on nuclear grade (low grade, intermediate grade and high grade) and intraductal necrosis (comedo or punctate). Imaging and clinical examination failed to identify the grade of DCIS due to lack of adequate resolution. Clinically, the histopathological gold standard "hematoxylin and eosin-stained (H and E) sections" is used to identify the grade of DCIS. But, hematoxylin and eosin-stained (H and E) sections has several shortcomings, including time-consuming and complicated pathological procedures, labor intensive, and some subjective errors. Multiphoton microscopy (MPM) based on two-photon excitation fluorescence (TPEF) and second harmonic generation (SHG), provides a label-free, low phototoxicity and high-resolution imaging method for label-free tissue samples. In this study, we used MPM to identify three different grades of human breast DCIS. Our results demonstrated that MPM can be rapidly used to identify three different grades of human breast DCIS on label-free tissue samples, via high-resolution imaging of intraductal proliferation of tumor cells, intraductal necrosis, and ductal basement membranes.
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
Pleomorphic invasive lobular carcinoma (pILC) is a distinct morphological variant of invasive lobular carcinoma (ILC), which is more aggressive than classic invasive lobular carcinoma (cILC) with worse overall prognosis. It is critical to distinguish between pILC and cILC for management and treatment of patients. Multiphoton microscopy (MPM),based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG), produces subcellular resolution images to show detailed information on structure and cell morphology of tissue sections without staining. In this work, MPM was introduced for label-free identification of normal breast lobules, pILC and cILC, which is consistent with haematoxylin and eosin (H&E) staining. Furthermore, analysis of nuclear area is used to quantitatively reflect their cytological characteristic. These results demonstrate that, MPM has the ability to distinguish between normal breast lobules, cILC, and pILC by histopathological features, which can provide additional indicators to determine adequate treatment strategies of patients with pILC for pathologists and surgeons. In the future, MPM may be used as a diagnostic tool of lobular cancer into routine clinical use.
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