van de Linde Sebastian, Heilemann Mike, Sauer Markus
Department of Biotechnology and Biophysics, Julius-Maximilians-University Würzburg, Germany.
Annu Rev Phys Chem. 2012;63:519-40. doi: 10.1146/annurev-physchem-032811-112012. Epub 2012 Jan 30.
Super-resolution imaging methods now can provide spatial resolution that is well below the diffraction limit approaching virtually molecular resolution. They can be applied to biological samples and provide new and exciting views on the structural organization of cells and the dynamics of biomolecular assemblies on wide timescales. These revolutionary developments come with novel requirements for fluorescent probes, labeling techniques, and data interpretation strategies. Synthetic fluorophores have a small size, are available in many colors spanning the whole spectrum, and can easily be chemically modified and used for stoichiometric labeling of proteins in live cells. Because of their brightness, their photostability, and their ability to be operated as photoswitchable fluorophores even in living cells under physiological conditions, synthetic fluorophores have the potential to substantially accelerate the broad application of live-cell super-resolution imaging methods.
超分辨率成像方法现在能够提供远低于衍射极限的空间分辨率,几乎接近分子分辨率。它们可应用于生物样本,并在宽时间尺度上为细胞的结构组织和生物分子组装体的动力学提供全新且令人兴奋的视角。这些革命性的进展对荧光探针、标记技术和数据解读策略提出了新的要求。合成荧光团尺寸小,有跨越整个光谱的多种颜色可供选择,并且能够轻松进行化学修饰,用于活细胞中蛋白质的化学计量标记。由于其亮度、光稳定性以及即使在生理条件下的活细胞中也能作为可光开关荧光团进行操作的能力,合成荧光团有潜力大幅加速活细胞超分辨率成像方法的广泛应用。