利用光活化有机荧光染料对固定和活细胞中的靶向蛋白进行超分辨率成像。
Superresolution imaging of targeted proteins in fixed and living cells using photoactivatable organic fluorophores.
机构信息
Department of Chemistry, Stanford University, Stanford, California 94305, USA.
出版信息
J Am Chem Soc. 2010 Nov 3;132(43):15099-101. doi: 10.1021/ja1044192.
Abstract
Superresolution imaging techniques based on sequential imaging of sparse subsets of single molecules require fluorophores whose emission can be photoactivated or photoswitched. Because typical organic fluorophores can emit significantly more photons than average fluorescent proteins, organic fluorophores have a potential advantage in super-resolution imaging schemes, but targeting to specific cellular proteins must be provided. We report the design and application of HaloTag-based target-specific azido DCDHFs, a class of photoactivatable push-pull fluorogens which produce bright fluorescent labels suitable for single-molecule superresolution imaging in live bacterial and fixed mammalian cells.
摘要
基于单分子稀疏子集的顺序成像的超分辨率成像技术需要荧光团，其发射可以光活化或光切换。因为典型的有机荧光团可以发射比平均荧光蛋白多得多的光子，所以有机荧光团在超分辨率成像方案中有潜在的优势，但必须提供针对特定细胞蛋白的靶向。我们报告了基于 HaloTag 的靶向叠氮 DCDHF 的设计和应用，这是一类光活化的推拉式荧光团，可产生适合活细菌和固定哺乳动物细胞中单分子超分辨率成像的明亮荧光标记。
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Nat Methods. 2010 Sep;7(9):717-9. doi: 10.1038/nmeth.1489. Epub 2010 Aug 8.
2
Molecules and methods for super-resolution imaging.用于超分辨率成像的分子和方法。
Methods Enzymol. 2010;475:27-59. doi: 10.1016/S0076-6879(10)75002-3.
3
Single-molecule and superresolution imaging in live bacteria cells.活细菌细胞中的单分子和超分辨率成像。
Cold Spring Harb Perspect Biol. 2010 Mar;2(3):a000448. doi: 10.1101/cshperspect.a000448.
4
Photoactivatable and photoconvertible fluorescent probes for protein labeling.光激活和光转化荧光探针用于蛋白质标记。
ACS Chem Biol. 2010 May 21;5(5):507-16. doi: 10.1021/cb1000229.
5
Single-molecule spectroscopy and imaging of biomolecules in living cells.活细胞中生物分子的单分子光谱和成像。
Anal Chem. 2010 Mar 15;82(6):2192-203. doi: 10.1021/ac9024889.
6
Caulobacter PopZ forms a polar subdomain dictating sequential changes in pole composition and function.钙杆状菌 PopZ 形成一个极性亚域，决定了极区组成和功能的顺序变化。
Mol Microbiol. 2010 Apr;76(1):173-89. doi: 10.1111/j.1365-2958.2010.07088.x. Epub 2010 Feb 10.
7
Why and how bacteria localize proteins.细菌为何以及如何定位蛋白质。
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