Department of Anatomy and Structural Biology, and.
Annu Rev Biophys. 2014;43:303-29. doi: 10.1146/annurev-biophys-051013-022836.
Superresolution fluorescence microscopy permits the study of biological processes at scales small enough to visualize fine subcellular structures that are unresolvable by traditional diffraction-limited light microscopy. Many superresolution techniques, including those applicable to live cell imaging, utilize genetically encoded photocontrollable fluorescent proteins. The fluorescence of these proteins can be controlled by light of specific wavelengths. In this review, we discuss the biochemical and photophysical properties of photocontrollable fluorescent proteins that are relevant to their use in superresolution microscopy. We then describe the recently developed photoactivatable, photoswitchable, and reversibly photoswitchable fluorescent proteins, and we detail their particular usefulness in single-molecule localization-based and nonlinear ensemble-based superresolution techniques. Finally, we discuss recent applications of photocontrollable proteins in superresolution imaging, as well as how these applications help to clarify properties of intracellular structures and processes that are relevant to cell and developmental biology, neuroscience, cancer biology and biomedicine.
超分辨率荧光显微镜允许在足够小的尺度上研究生物过程，从而可以可视化传统衍射受限光显微镜无法分辨的精细亚细胞结构。许多超分辨率技术，包括适用于活细胞成像的技术，都利用遗传编码的光控可激活荧光蛋白。这些蛋白质的荧光可以通过特定波长的光来控制。在这篇综述中，我们讨论了光控可激活荧光蛋白的生化和光物理特性，这些特性与其在超分辨率显微镜中的应用相关。然后，我们描述了最近开发的光激活型、光转换型和可逆光转换型荧光蛋白，并详细说明了它们在基于单分子定位和非线性整体的超分辨率技术中的特殊用途。最后，我们讨论了光控可激活蛋白在超分辨率成像中的最新应用，以及这些应用如何帮助阐明与细胞和发育生物学、神经科学、癌症生物学和生物医学相关的细胞内结构和过程的特性。
