Diabetic Nephropathy - Methods and Protocols
Diabetic Nephropathy - Methods and Protocols
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糖尿病肾病 - 方法和方案
DOI：
10.1007/978-1-4939-9841-8_8
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Jafree D
中科院分区：
文献类型：
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作者：
Jafree D
三维生物组织的微观和宏观评价正变得越来越流行。这一趋势与众多组织清除策略的出现以及共聚焦和双光子显微镜的进步相一致，使完整器官和系统的研究能够达到细胞和亚细胞分辨率。在这一章中，我们描述了一个全装载免疫荧光技术标记结构在肾组织。该技术结合溶剂型组织透明化和共聚焦成像，无论是否使用双光子激发，都能提供比传统切片和单独染色更丰富的结构信息。鉴于我们的方法中加入了石蜡包埋，这种混合方案提供了一种强有力的方法，将联合收割机共聚焦或双光子结果与组织学和同一组织内的进一步免疫荧光分析相结合。
Microscopic and macroscopic evaluation of biological tissues in three dimensions is becoming increasingly popular. This trend is coincident with the emergence of numerous tissue clearing strategies, and advancements in confocal and two-photon microscopy, enabling the study of intact organs and systems down to cellular and sub-cellular resolution. In this chapter, we describe a wholemount immunofluorescence technique for labeling structures in renal tissue. This technique combined with solvent-based tissue clearing and confocal imaging, with or without two-photon excitation, provides greater structural information than traditional sectioning and staining alone. Given the addition of paraffin embedding to our method, this hybrid protocol offers a powerful approach to combine confocal or two-photon findings with histological and further immunofluorescent analysis within the same tissue.