Neural stem cells (NSCs) are self-renewing, multipotent cells which give rise to all components of the central nervous system (CNS) during embryogenesis, but also activate in response to injury and disease and maintain a certain level of neurogenic activity throughout adulthood. This activity takes place in specialized regions of the brain, the neurovascular niches, whose main role is to control the behaviour of the CNS. In adult mammals, two main “canonical” niches have been described: The subventricular zone (SVZ) of the lateral ventricles and the subgranular zone (SGZ) of the dentate gyrus. This review discusses our current understanding of the neural stem cells and their canonical niches, as well as their structure, behaviours, and role in neural disease.
Cutaneous melanoma is one of the most aggressive and deadliest cancers in human beings due to its invasiveness and other factors. Histopathological analysis is crucial for a proper diagnosis. Optical tissue clearing is a novel field that allows 3D image acquisition of large-scale biological tissues. Optical clearing and immunolabeling for 3D fluorescence imaging has yet to be extensively applied to melanoma. In the present manuscript, we establish, for the first time, an optical clearing and immunostaining procedure for human melanoma and human cell line-derived melanoma xenograft models using the CUBIC (clear, unobstructed brain imaging cocktails) technique. We have successfully cleared the samples and achieved 3D volumetric visualization of the tumor microenvironment, vasculature, and cell populations.
: Optical tissue clearing techniques can render large-scale tissue samples and whole organs transparent to apply cell-resolution 3D imaging techniques, revolutionizing the fields of biological research and pathological diagnosis, and providing never-before seen structural information. There are several widely used protocols, but they have shortcomings, both in complexity of use and in suboptimal results in certain tissues and organs. Here, we describe a new method dubbed “White and Clear” (WaC) using 2,2’-thiodiethanol (TDE) as a dynamic Refractive Index (RI)-matching reagent alongside a lipid-extraction and a pigment-bleaching step using dilute hydrogen peroxide (H2O2), to provide ease of use and enhanced clearing effect on all kinds of samples and allow for the clearing parameters to be finely adjusted in order to obtain the best results in each situation.