Shroff Urvi Nikhil, Schiessl Ina Maria, Gyarmati Georgina, Riquier-Brison Anne, Peti-Peterdi Janos
Department of Physiology and Neuroscience, Zilkha Neurogenetic Institute, Keck School of Medicine, University of Southern California, Los Angeles, CA, United States.
Department of Physiology and Neuroscience, Zilkha Neurogenetic Institute, Keck School of Medicine, University of Southern California, Los Angeles, CA, United States.
Methods Cell Biol. 2019;154:85-107. doi: 10.1016/bs.mcb.2019.04.013. Epub 2019 May 17.
Fluorescence microscopy techniques are powerful tools to study tissue dynamics, cellular function and biology both in vivo and in vitro. These tools allow for functional assessment and quantification along with qualitative analysis, thus providing a comprehensive understanding of various cellular processes under normal physiological and disease conditions. The main focus of this chapter is the recently developed method of serial intravital multiphoton microscopy that has helped shed light on the dynamic alterations of the spatial distribution and fate of single renal cells or cell populations and their migration patterns in the same tissue region over several days in response to various stimuli within the living kidney. This technique is very useful for studying in vivo the molecular and cellular mechanisms of tissue remodeling and repair after injury. In addition, complementary in vitro imaging tools are also described and discussed, like tissue clearing techniques and protein synthesis measurement in tissues in situ that provide an in depth assessment of changes at the cellular level. Thus, these novel fluorescence techniques can be effectively leveraged for different tissue types, experimental conditions as well as disease models to improve our understanding of renal cell biology.
荧光显微镜技术是研究体内和体外组织动态、细胞功能及生物学特性的强大工具。这些工具不仅能进行定性分析,还能实现功能评估和定量分析,从而全面了解正常生理和疾病状态下的各种细胞过程。本章的主要重点是最近开发的连续活体多光子显微镜方法,该方法有助于揭示单个肾细胞或细胞群体在活体肾脏中,在几天内对各种刺激做出反应时,其空间分布、命运及迁移模式的动态变化。这项技术对于研究损伤后组织重塑和修复的体内分子和细胞机制非常有用。此外,还介绍并讨论了互补的体外成像工具,如组织透明化技术和组织原位蛋白质合成测量,这些工具能深入评估细胞水平的变化。因此,这些新型荧光技术可有效地应用于不同组织类型、实验条件及疾病模型,以增进我们对肾细胞生物学的理解。