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Ratiometric electrochemiluminescence sensing and intracellular imaging of ClO− via resonance energy transfer

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Abstract

Electrochemiluminescence resonance energy transfer (ECL-RET) is a versatile signal transduction strategy widely used in the fabrication of chem/biosensors. However, this technique has not yet been applied in visualized imaging analysis of intracellular species due to the insulating nature of the cell membrane. Here, we construct a ratiometric ECL-RET analytical method for hypochlorite ions (ClO−) by ECL luminophore, with a luminol derivative (L-012) as the donor and a fluorescence probe (fluorescein hydrazide) as the acceptor. L-012 can emit a strong blue ECL signal and fluorescein hydrazide has negligible absorbance and fluorescence signal in the absence of ClO−. Thus, the ECL-RET process is turned off at this time. In the presence of ClO−, however, the closed-loop hydrazide structure in fluorescein hydrazide is opened via specific recognition with ClO−, accompanied with intensified absorbance and fluorescence signal. Thanks to the spectral overlap between the ECL spectrum of L-012 and the absorption spectrum of fluorescein, the ECL-RET effect is gradually recovered with the addition of ClO−. Furthermore, the ECL-RET system has been successfully applied to image intracellular ClO−. Although the insulating nature of the cell itself can generate a shadow ECL pattern in the cellular region, extracellular ECL emission penetrates the cell membrane and excites intracellular fluorescein generated by the reactions between fluorescein hydrazide and ClO−. The cell imaging strategy via ECL-RET circumvents the blocking of the cell membrane and enables assays of intracellular species. The importance of the ECL-RET platform lies in calibrating the fluctuation from the external environment and improving the selectivity by using fluorescent probes. Therefore, this ratiometric ECL sensor has shown broad application prospects in the identification of targets in clinical diagnosis and environmental monitoring.

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Funding

This work was supported by the National Natural Science Foundation of China grants 22374125, 22201244, and 22076161; China Postdoctoral Science Foundation (2022M722688); Yangzhou University Interdisciplinary Research Foundation for Chemistry Discipline of Targeted Support yzuxk202009; Foundation of State Key Laboratory of Analytical Chemistry for Life Science grant No. SKLACLS2201; Lvyangjinfeng Talent Program of Yangzhou; Talent Support Program of Yangzhou University; The Open Project Program of Jiangsu Key Laboratory of Zoonosis (No. R2013); and The project funded by the PAPD and TAPP.

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  1. School of Chemistry and Chemical Engineering, Yangzhou University, YangzhouJiangsu, 225002, China

    Cheng Ma, Yujing Zhu, Zhichen Zhang, Xuan Chen, Zhengping Ji, Lu-Nan Zhang & Qin Xu

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  1. Cheng Ma
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  2. Yujing Zhu
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  3. Zhichen Zhang
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  4. Xuan Chen
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Contributions

C.M. and Q.X. conceived the study. Y.Z., C.M., and Z.Z. performed the experiments. C.M. and Y.Z. built the microscope setup. X.C., Z.J., L.N.Z., and C.M. advised the manuscript. C.M. and Q.X. designed the experiments. Y.Z. and C.M. analyzed the data. Y.Z., C.M., and Q.X. wrote the paper.

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Correspondence to Cheng Ma or Qin Xu.

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Published in the topical collection Emerging Trends in Electrochemical Analysis with guest editors Sabine Szunerits, Wei Wang, and Adam T. Woolley.

Cheng Ma and Yujing Zhu contributed equally to this work.

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Ma, C., Zhu, Y., Zhang, Z. et al. Ratiometric electrochemiluminescence sensing and intracellular imaging of ClO− via resonance energy transfer. Anal Bioanal Chem (2024). https://doi.org/10.1007/s00216-024-05236-6

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