Superresolution imaging of targeted proteins in fixed and living cells using photoactivatable organic fluorophores

Hsiao Lu D. Lee; Samuel J. Lord; Shigeki Iwanaga; Ke Zhan; Hexin Xie; Jarrod C. Williams; Hui Wang; Grant R. Bowman; Erin D. Goley; Lucy Shapiro; Robert J. Twieg; Jianghong Rao; W. E. Moerner

doi:10.1021/ja1044192

Superresolution imaging of targeted proteins in fixed and living cells using photoactivatable organic fluorophores

Hsiao Lu D. Lee, Samuel J. Lord, Shigeki Iwanaga, Ke Zhan, Hexin Xie, Jarrod C. Williams, Hui Wang, Grant R. Bowman, Erin D. Goley, Lucy Shapiro, Robert J. Twieg, Jianghong Rao, W. E. Moerner

Research output: Contribution to journal › Article › peer-review

133 Scopus citations

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.

Original language	English (US)
Pages (from-to)	15099-15101
Number of pages	3
Journal	Journal of the American Chemical Society
Volume	132
Issue number	43
DOIs	https://doi.org/10.1021/ja1044192
State	Published - Nov 3 2010
Externally published	Yes

ASJC Scopus subject areas

Catalysis
Chemistry(all)
Biochemistry
Colloid and Surface Chemistry

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10.1021/ja1044192

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Lee, H. L. D., Lord, S. J., Iwanaga, S., Zhan, K., Xie, H., Williams, J. C., Wang, H., Bowman, G. R., Goley, E. D., Shapiro, L., Twieg, R. J., Rao, J., & Moerner, W. E. (2010). Superresolution imaging of targeted proteins in fixed and living cells using photoactivatable organic fluorophores. Journal of the American Chemical Society, 132(43), 15099-15101. https://doi.org/10.1021/ja1044192

Lee, HLD, Lord, SJ, Iwanaga, S, Zhan, K, Xie, H, Williams, JC, Wang, H, Bowman, GR, Goley, ED, Shapiro, L, Twieg, RJ, Rao, J & Moerner, WE 2010, 'Superresolution imaging of targeted proteins in fixed and living cells using photoactivatable organic fluorophores', Journal of the American Chemical Society, vol. 132, no. 43, pp. 15099-15101. https://doi.org/10.1021/ja1044192

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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.",

author = "Lee, {Hsiao Lu D.} and Lord, {Samuel J.} and Shigeki Iwanaga and Ke Zhan and Hexin Xie and Williams, {Jarrod C.} and Hui Wang and Bowman, {Grant R.} and Goley, {Erin D.} and Lucy Shapiro and Twieg, {Robert J.} and Jianghong Rao and Moerner, {W. E.}",

year = "2010",

month = nov,

day = "3",

doi = "10.1021/ja1044192",

language = "English (US)",

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T1 - Superresolution imaging of targeted proteins in fixed and living cells using photoactivatable organic fluorophores

AU - Lee, Hsiao Lu D.

AU - Lord, Samuel J.

AU - Iwanaga, Shigeki

AU - Zhan, Ke

AU - Xie, Hexin

AU - Williams, Jarrod C.

AU - Wang, Hui

AU - Bowman, Grant R.

AU - Goley, Erin D.

AU - Shapiro, Lucy

AU - Twieg, Robert J.

AU - Rao, Jianghong

AU - Moerner, W. E.

PY - 2010/11/3

Y1 - 2010/11/3

N2 - 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.

AB - 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.

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Superresolution imaging of targeted proteins in fixed and living cells using photoactivatable organic fluorophores

Abstract

ASJC Scopus subject areas

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