Glioblastoma (GBM) is a highly malignant and rapidly invasive astrocytoma, which has explosive biologic properties with rapid clinical progression leading to death and has a poor clinical outcome. The average survival time of most patients is only 12 to 15 months. GBM is distinguished pathologically from lower grade tumors by ‘pseudopalisading’ necrosis and microvascular hyperplasia. The most exaggerated form of microvascular hyperplasia is called glomeruloid body. MPM is a potential tool for imaging biological tissues at the molecular level. In this paper, MPM based on twophoton excited fluorescence (TPEF) and second harmonic generation (SHG) was applied for identifying the GBM without labeling or fluorescent markers. The results showed that MPM can display the specific histological characteristics of GBM including ‘pseudopalisading’ necrosis and glomeruloid vascular proliferation. The results obtained are consistent with the diagnosis of pathological findings. MPM will become a promising imaging tool for preoperative diagnosis of glioblastoma in the future.
Chronic obstructive pulmonary diseases (COPD) is the fifth leading cause of death worldwide and will be increased in the coming decades. Pulmonary emphysema is one of the hallmarks of COPD. Establishing the animal model of pulmonary emphysema is very important to explore its pathogenesis. Until now, researchers are still having used histological methods to assess whether it is successful to stimulate the emphysema animal model. In this study, we try to use multiphoton microscopy imaging system to assess whether the mouse models obtained the emphysema pathological. The two-photon excited fluorescence (TPEF) signals and second harmonic generation (SHG) signals clearly showed changes in both cellular features and extracellular matrix architecture during different time of emphysema mouse models. With the development of miniaturized multiphoton microscopy, multiphoton microscopy can be used to monitor the developing of pulmonary emphysema in animals in vivo.
Cortical structures in the central nervous system exhibit an ordered laminar organization. Defined cell layers are significant to our understanding of brain structure and function. In this work, multiphoton microscopy (MPM) based on second harmonic generation (SHG) and two-photon excited fluorescence (TPEF), which was applied for qualitatively visualizing the structure of cerebral and cerebellar cortex from the fresh, unfixed, and unstained specimen. MPM is able to effectively identify neurons and neurites in cerebral cortex, as well as glial cells, Purkinje cells, and granule cells in cerebellar cortex at subcellular resolution. In addition, the use of automated image processing algorithms can quantify the circularity of neurons and the density distribution of neurites based on the intrinsic nonlinear optical contrast, further providing quantitative characteristics for automatically analyzing the laminar structure of cortical structures. These results suggest that with the development of the feasibility of two-photon fiberscopes and microendoscope probes, the combined MPM and image analysis holds potential to provide supplementary information to augment the diagnostic accuracy of neuropathology and in vivo identification of various neurological illnesses in clinic.