Heart Research Center Goettingen, University Medicine Goettingen, Germany.
J Mol Cell Cardiol. 2013 May;58:13-21. doi: 10.1016/j.yjmcc.2012.11.016. Epub 2012 Dec 3.
Detailed understanding of the adaptive nature of cardiac cells in health and disease requires investigation of proteins and membranes in their native physiological environment, ideally by noninvasive optical methods. However, conventional light microscopy does not resolve the spatial characteristics of small fluorescently labeled protein or membrane structures in cells. Due to diffraction limiting resolution to half the wavelength of light, adjacent fluorescent molecules spaced at less than ~250 nm are not separately visualized. This fundamental problem has lead to a rapidly growing area of research, superresolution fluorescence microscopy, also called nanoscopy. We discuss pioneering applications of superresolution microscopy relevant to the heart, emphasizing different nanoscopy strategies toward new insight in cardiac cell biology. Here, we focus on molecular and structural readouts from subcellular nanodomains and organelles related to Ca(2+) signaling during excitation-contraction (EC) coupling, including live cell imaging strategies. Based on existing data and superresolution techniques, we suggest that an important future aim will be subcellular in situ structure-function analysis with nanometric resolving power in organotypic cells.
详细了解心脏细胞在健康和疾病中的适应特性需要在其天然生理环境中研究蛋白质和膜，理想情况下是通过非侵入性的光学方法。然而，传统的光学显微镜无法解析细胞中小荧光标记蛋白或膜结构的空间特征。由于衍射限制了分辨率至光波长的一半，因此小于约 250nm 的相邻荧光分子不能被单独可视化。这个基本问题已经导致了一个快速发展的研究领域，即超分辨率荧光显微镜，也称为纳米显微镜。我们讨论了与心脏相关的超分辨率显微镜的开创性应用，强调了不同的纳米显微镜策略在心脏细胞生物学中的新见解。在这里，我们重点介绍与兴奋-收缩（EC）偶联期间 Ca(2+)信号相关的亚细胞纳米域和细胞器的分子和结构读出，包括活细胞成像策略。基于现有数据和超分辨率技术，我们认为一个重要的未来目标将是在器官型细胞中具有纳米级分辨率的亚细胞原位结构-功能分析。
