Organic solvent‐based tissue clearing techniques and their applications

Zhan, Yanjing; Wu, Haoyan; Liu, Linfeng; Lin, Jie; Zhang, Shiwen

doi:10.1002/jbio.202000413

Cited by 19 publications

(14 citation statements)

References 76 publications

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“…Besides, tissue-clearance technique is applied in our lab. 224 , 225 For example, we are able to visualize the bone-implant interfaces after tissue-clearance with two-photon excitation microscope, which is almost unapplicable with other methods.…”

Section: Disputes and Challenges Of Sscsmentioning

confidence: 99%

Insights into skeletal stem cells

Lei

et al. 2022

Bone Res

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The tissue-resident skeletal stem cells (SSCs), which are self-renewal and multipotent, continuously provide cells (including chondrocytes, bone cells, marrow adipocytes, and stromal cells) for the development and homeostasis of the skeletal system. In recent decade, utilizing fluorescence-activated cell sorting, lineage tracing, and single-cell sequencing, studies have identified various types of SSCs, plotted the lineage commitment trajectory, and partially revealed their properties under physiological and pathological conditions. In this review, we retrospect to SSCs identification and functional studies. We discuss the principles and approaches to identify bona fide SSCs, highlighting pioneering findings that plot the lineage atlas of SSCs. The roles of SSCs and progenitors in long bone, craniofacial tissues, and periosteum are systematically discussed. We further focus on disputes and challenges in SSC research.

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Section: Disputes and Challenges Of Sscsmentioning

confidence: 99%

Insights into skeletal stem cells

Lei

et al. 2022

Bone Res

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“…Clearing methods for preserving tissue morphology are principally divided into three categories: solvent‐based methods (for a review, see Zhan, Wu, Liu, Lin, & Zhang, 2021) aqueous‐based methods (Tainaka et al, 2018) hydrogel‐based methods (Choi, Guan, & Chung, 2021; Flood, Page, & Reynaud, 2016; Gradinaru, Treweek, Overton, & Deisseroth, 2018) …”

Section: Clearing Fixed Samplesmentioning

confidence: 99%

Lightsheet Microscopy

et al. 2022

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In this paper, we review lightsheet (selective plane illumination) microscopy for mouse developmental biologists. There are different means of forming the illumination sheet, and we discuss these. We explain how we introduced the lightsheet microscope economically into our core facility and present our results on fixed and living samples. We also describe methods of clearing fixed samples for three-dimensional imaging and discuss the various means of preparing samples with particular reference to mouse cilia, adipose spheroids, and cochleae.

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“…Sample size and tissue composition impact the clearing speed and limit the microscopes that can be used when imaging large samples (21). Fluorophore preservation poses another challenge as certain clearing protocols are incompatible with fluorescent probes (22). In particular, preservation of protein-based fluorescence and lipid staining remain open challenges in the field (16).…”

Section: Introductionmentioning

confidence: 99%

Comparison between optical tissue clearing methods for detecting administered mesenchymal stromal cells in mouse lungs

Pichardo

Amadeo

Wilm

et al. 2022

Preprint

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Optical tissue clearing of lung tissue enables the intact lung to be imaged using fluorescence microscopy. Several clearing protocols have been developed in recent years, including the Clear, Unobstructed Brain/Body Imaging Cocktails and Computational analysis (CUBIC), stabilised 3D imaging of solvent-cleared organs (s-DISCO) and Ethyl cinnamate (ECi) methods. Here we compared these protocols with the aim of determining the biodistribution of mesenchymal stroma cells (MSCs) and understanding how they interact with host cells in the mouse lung. First, we evaluated how each method affected the size, morphology, and transparency of the lungs. Then, we compared the preservation of the fluorescence of the protein tdTomato expressed by the MSCs, and of the organic dye Evans Blue which labels the vasculature. In addition, we tested the compatibility of the methods with immunofluorescence staining. We found that CUBIC clearing is the only method that enables direct imaging of fluorescently labelled MSCs in the lungs thereby allowing the study of the MSC interaction with endothelial and immune cells when combined with immunofluorescence staining. Overall, 3D imaging of CUBIC cleared lungs confirmed that injected MSCs are initially retained in the pulmonary microvasculature, and that most cells are eliminated from the lungs within the first 24 h.

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Organic solvent‐based tissue clearing techniques and their applications

Cited by 19 publications

References 76 publications

Insights into skeletal stem cells

Insights into skeletal stem cells

Lightsheet Microscopy

Comparison between optical tissue clearing methods for detecting administered mesenchymal stromal cells in mouse lungs

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