Dunn Kenneth W, Sutton Timothy A, Sandoval Ruben M
Indiana University School of Medicine, Indianapolis, Indiana.
Curr Protoc Cytom. 2018 Jan 18;83:12.9.1-12.9.25. doi: 10.1002/cpcy.32.
Intravital microscopy, microscopy of living animals, is a powerful research technique that combines the resolution and sensitivity found in microscopic studies of cultured cells with the relevance and systemic influences of cells in the context of the intact animal. The power of intravital microscopy has recently been extended with the development of multiphoton fluorescence microscopy systems capable of collecting optical sections from deep within the kidney at subcellular resolution, supporting high-resolution characterizations of the structure and function of glomeruli, tubules, and vasculature in the living kidney. Fluorescent probes are administered to an anesthetized, surgically prepared animal, followed by image acquisition for up to 3 hr. Images are transferred via a high-speed network to specialized computer systems for digital image analysis. This general approach can be used with different combinations of fluorescent probes to evaluate processes such as glomerular permeability, proximal tubule endocytosis, microvascular flow, vascular permeability, mitochondrial function, and cellular apoptosis/necrosis. © 2018 by John Wiley & Sons, Inc.
活体显微镜检查，即对活体动物进行显微镜检查，是一种强大的研究技术，它将培养细胞微观研究中的分辨率和灵敏度与完整动物体内细胞的相关性及系统性影响相结合。随着多光子荧光显微镜系统的发展，活体显微镜检查的能力最近得到了扩展，该系统能够以亚细胞分辨率从肾脏深处采集光学切片，支持对活体肾脏中肾小球、肾小管和脉管系统的结构和功能进行高分辨率表征。将荧光探针给予麻醉且经过手术准备的动物，随后进行长达3小时的图像采集。图像通过高速网络传输到专门的计算机系统进行数字图像分析。这种通用方法可与不同的荧光探针组合使用，以评估诸如肾小球通透性、近端肾小管内吞作用、微血管血流、血管通透性、线粒体功能以及细胞凋亡/坏死等过程。© 2018约翰威立国际出版公司
