Browsing by Subject "Tissue clearing"

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    Open Access
    A qualitative comparison of ten tissue clearing techniques
    (Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Orlich, Michael; Kiefer, Friedemann
    The understanding of spatially complex biological systems is greatly aided by the availability of high resolution information on their tissue architecture, as is provided by optical sectioning microscopy like confocal and light sheet microscopy. In addition, genetically encoded fluorescent reporter proteins reveal tissue architecture without the need for staining procedures. Owing to opacity caused by scattering and absorption, light microscopy in tissue is limited to thin tissue layers of a few micrometers traditionally provided by histological sections. Aiming to allow deeper imaging, during the last decade massive efforts to develop tissue clearing protocols produced a flurry of novel clearing techniques for whole organ visualization, now available to microscopists. In particular, new tissue clearing methods were developed that avoid the use of organic solvents, aiming to retain the integrity of genetically encoded fluorescent proteins. So far, these methods have not been directly compared and selection of the right technique can be a non-trivial task. Here, we have aimed to compare different tissue clearing approaches side by side in a standardized manner. We provide qualitative data on their clearing capability of mouse brain, lung, heart, kidney and muscle, as well as embryos and fetuses at the developmental stages E10.5, E12.5 and E15.5 and discuss possible applications.
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    Publication
    Open Access
    Progress on three-dimensional visualizing skin architecture with multiple immunofluorescence staining and tissue-clearing approaches
    (Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2025) Wang, Yuqing; Bai, Wanzhu; Wang, Xiaoyu
    The skin forms the external covering of the body and is its largest organ, comprising many different cell types. Although the diversity of these cells has been widely studied with various histological methods, our understanding of skin architecture is mainly established on thin tissue sections, which restricted the information available to two dimensions. The development of innovative techniques to induce optical transparency (“clearing”) in biological tissues has enabled researchers to visualize the three-dimensional reconstruction of intact organs and thick tissue sections at a cellular resolution. With the aid of tissue-clearing treatment, the labeled cutaneous nerve fibers and blood vessels can be followed for a longer distance on the thicker skin section or the whole mount skin under a fluorescence microscopy or a confocal microscopy. It is beneficial for demonstrating the morphological characteristics of nerve fibers and blood vessels themselves, as well as their spatial interconnection. In this review, we provide a brief summary of the literature on the use of tissue optical clearing methods and describe our experience of multiple fluorescent staining and tissue clearing approaches on thicker skin sections and whole-mount skin in our laboratory. Given the existing conventional methods, we expected to provide a more effective approach to comprehensively study skin architecture.