ESIPT-Based Photoactivatable Fluorescent Probe for Ratiometric Spatiotemporal Bioimaging

doi:10.3390/s16101684

. 2016 Oct 12;16(10):1684.

doi: 10.3390/s16101684.

ESIPT-Based Photoactivatable Fluorescent Probe for Ratiometric Spatiotemporal Bioimaging

Xiaohong Zhou^{1

2}, Yuren Jiang³, Xiongjie Zhao⁴, Dong Guo⁵

Affiliations

¹ College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China. zhouxh001@csu.edu.cn.
² Environment Monitoring Department, Changsha Environmental Protection College, Changsha 410004, China. zhouxh001@csu.edu.cn.
³ College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China. jiangyr@mail.csu.edu.cn.
⁴ College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China. 152301025@csu.edu.cn.
⁵ College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China. guo4675@csu.edu.cn.

PMID: 27754338
PMCID: PMC5087472
DOI: 10.3390/s16101684

ESIPT-Based Photoactivatable Fluorescent Probe for Ratiometric Spatiotemporal Bioimaging

Xiaohong Zhou et al. Sensors (Basel). 2016.

. 2016 Oct 12;16(10):1684.

doi: 10.3390/s16101684.

Authors

Xiaohong Zhou^{1

2}, Yuren Jiang³, Xiongjie Zhao⁴, Dong Guo⁵

Affiliations

¹ College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China. zhouxh001@csu.edu.cn.
² Environment Monitoring Department, Changsha Environmental Protection College, Changsha 410004, China. zhouxh001@csu.edu.cn.
³ College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China. jiangyr@mail.csu.edu.cn.
⁴ College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China. 152301025@csu.edu.cn.
⁵ College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China. guo4675@csu.edu.cn.

PMID: 27754338
PMCID: PMC5087472
DOI: 10.3390/s16101684

Abstract

Photoactivatable fluorophores have become an important technique for the high spatiotemporal resolution of biological imaging. Here, we developed a novel photoactivatable probe (PHBT), which is based on 2-(2-hydroxyphenyl)benzothiazole (HBT), a small organic fluorophore known for its classic luminescence mechanism through excited-state intramolecular proton transfer (ESIPT) with the keto form and the enol form. After photocleavage, PHBT released a ratiometric fluorophore HBT, which showed dual emission bands with more than 73-fold fluorescence enhancement at 512 nm in buffer and more than 69-fold enhancement at 452 nm in bovine serum. The probe displayed a high ratiometric imaging resolution and is believed to have a wide application in biological imaging.

Keywords: ESIPT; photoactivatable; ratiometric.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

**Scheme 1**
Synthetic route for the photoactivatable fluorescent probe PHBT and the response mechanism. (a) R: H₂O₂ (30%), R: (NH₄)₂Ce(NO₃)₆, S: MeCN, 30 min, room temperature; (b) R: K₂CO₃, S: MeCN, 24 h, 70 °C.

**Figure 1**
(a,b) Time-dependent fluorescent spectral changes of PHBT under UV irradiation (0–30 min) at room temperature, PHBT dissolved in PBS buffer (10 mM, pH 7.4, water/DMSO = 99:1) to form 10 μM testing solution, with excitation λ = 365 nm. (inset) Change in the fluorescence of the probe (**left**) before and (**right**) after UV irradiation (30 min) at room temperature.

**Figure 2**
(a,b) Time-dependent fluorescent spectral changes of PHBT under UV irradiation (0–30 min) at room temperature, PHBT dissolved in PBS buffer (10 mM, pH 7.4, aqueous 20% bovine serum solution) to form 10 μM testing solution with excitation λ = 365 nm.

**Figure 3**
Fluorescent images. MDA-MB-231 cells stained with 10 µM PHBT. (a) Bright field image of PHBT in cells; (b) green fluorescence of PHBT in cells after 30 min of irradiation at 405 nm; (c) red fluorescence of PHBT in cells; and (d) overlay of (b,c).

See this image and copyright information in PMC

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[1] Bergauer B., Knipfer C., Amann A., Rohde M., Tangermann-Gerk K., Adler W., Schmidt M., Nkenke E., Stelzle F. Does laser surgery interfere with optical nerve identification in maxillofacial hard and soft tissue?—An experimental ex vivo study. Sensors. 2015;15:25416–25432. doi: 10.3390/s151025416. - DOI - PMC - PubMed

[2] Bergauer B., Knipfer C., Amann A., Rohde M., Tangermann-Gerk K., Adler W., Schmidt M., Nkenke E., Stelzle F. Does laser surgery interfere with optical nerve identification in maxillofacial hard and soft tissue?—An experimental ex vivo study. Sensors. 2015;15:25416–25432. doi: 10.3390/s151025416. - DOI - PMC - PubMed

[3] Lim R.K.V., Lin Q. Photoinducible bioorthogonal chemistry: A spatiotemporally controllable tool to visualize and perturb proteins in live cells. Acc. Chem. Res. 2011;44:828–839. doi: 10.1021/ar200021p. - DOI - PMC - PubMed

[4] Lim R.K.V., Lin Q. Photoinducible bioorthogonal chemistry: A spatiotemporally controllable tool to visualize and perturb proteins in live cells. Acc. Chem. Res. 2011;44:828–839. doi: 10.1021/ar200021p. - DOI - PMC - PubMed

[5] Vaughan J.C., Jia S., Zhuang X. Ultrabright photoactivatable fluorophores created by reductive caging. Nat. Methods. 2012;9:1181–1184. doi: 10.1038/nmeth.2214. - DOI - PMC - PubMed

[6] Vaughan J.C., Jia S., Zhuang X. Ultrabright photoactivatable fluorophores created by reductive caging. Nat. Methods. 2012;9:1181–1184. doi: 10.1038/nmeth.2214. - DOI - PMC - PubMed

[7] Vaillancourt R.R., Dhanasekaran N., Johnson G.L., Ruoho A.E. 2-Azido-[32P]NAD+, a photoactivatable probe for G-protein structure: Evidence for holotransducin oligomers in which the ADP-ribosylated carboxyl terminus of a interacts with both a and y subunits. Proc. Natl. Acad. Sci. USA. 1990;87:3645–3649. doi: 10.1073/pnas.87.10.3645. - DOI - PMC - PubMed

[8] Vaillancourt R.R., Dhanasekaran N., Johnson G.L., Ruoho A.E. 2-Azido-[32P]NAD+, a photoactivatable probe for G-protein structure: Evidence for holotransducin oligomers in which the ADP-ribosylated carboxyl terminus of a interacts with both a and y subunits. Proc. Natl. Acad. Sci. USA. 1990;87:3645–3649. doi: 10.1073/pnas.87.10.3645. - DOI - PMC - PubMed

[9] Lee S.H., Shin J.Y., Lee A.C. Counting single photoactivatable fluorescent molecules by photoactivated localization microscopy (PALM) Bustamante. 2012;109:17436–17441. doi: 10.1073/pnas.1215175109. - DOI - PMC - PubMed

[10] Lee S.H., Shin J.Y., Lee A.C. Counting single photoactivatable fluorescent molecules by photoactivated localization microscopy (PALM) Bustamante. 2012;109:17436–17441. doi: 10.1073/pnas.1215175109. - DOI - PMC - PubMed

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ESIPT-Based Photoactivatable Fluorescent Probe for Ratiometric Spatiotemporal Bioimaging

Affiliations

ESIPT-Based Photoactivatable Fluorescent Probe for Ratiometric Spatiotemporal Bioimaging

Authors

Affiliations

Abstract

Conflict of interest statement

Figures

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References

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