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
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.
Standard histopathology is well accepted as the gold standard for the diagnosis a wide range of diseases. Despite continuing advances in tissue staining automation, typical histological processing such as formalin-fixed paraffin-embedded are also labour- and time-intensive for treatment decisions in intraoperative histopathologic diagnosis. Multiphoton microscopy (MPM), based on second harmonic generation (SHG) and two-photon excited fluorescence (TPEF), can be a versatile tool that enables label-free mapping of endogenous fluorophores within a fresh specimen, which provides pathology-like images with cellular and subcellular details. Here, we describe the use of label-free MPM for visualizing rat and human ex vivo brain tissue without tissue fixation, processing, and staining. Moreover, MPM is able to identify 6 types of cells in rat cerebrum and cerebellum, including cortical neurons, glia cells, Purkinje cells, pyramidal neurons and granule neurons in hippocampus, as well as epithelial cells in lateral ventricle. In addition, we further demonstrate that MPM can provide definitive pathological features in cerebral ischemia and focal cortical dysplasia (FCD) for assisting pathologic diagnosis. Our work establishes the methodology and augments the diagnostic accuracy of traditional frozen section histopathology. With the development of the miniature two-photon microscope, MPM will show more potential as a practical clinical tool for providing intraoperative reference image guidance of resection in neurosurgery.