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Photocontrollable fluorescent proteins for superresolution imaging.
Annual Review of Biophysics ( IF 10.4 ) Pub Date : 2014-01-01 , DOI: 10.1146/annurev-biophys-051013-022836
Daria M Shcherbakova 1 , Prabuddha Sengupta , Jennifer Lippincott-Schwartz , Vladislav V Verkhusha
Affiliation  

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.

中文翻译:

用于超分辨率成像的光控荧光蛋白。

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,以可视化传统衍射极限光学显微镜无法解析的精细亚细胞结构。许多超分辨率技术,包括那些适用于活细胞成像的技术,利用遗传编码的光控荧光蛋白。这些蛋白质的荧光可以由特定波长的光控制。在这篇综述中,我们讨论了与它们在超分辨率显微镜中的使用相关的光控荧光蛋白的生化和光物理特性。然后,我们描述了最近开发的可光活化、光开关和可逆光开关荧光蛋白,我们详细介绍了它们在基于单分子定位和基于非线性集合的超分辨率技术中的特殊用途。最后,我们讨论了光控蛋白质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。
更新日期:2014-06-04
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