Journal
TALANTA
Volume 220, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.talanta.2020.121355
Keywords
Intracellular Cu2+; Intracellular uptaken copper; Fluorescence imaging; Inductively coupled plasma mass spectrometry; Quantification
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Funding
- National Natural Science Foundation of China [21974018, 21727811]
- Fundamental Research Funds for the Central Universities of China [N2005015, N2005027]
- Liaoning Revitalization Talents Program [XLYC1907191, XLYC1802016]
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A variety of fluorescence probes have been developed for fluorescence imaging of metals in biological cells. However, accurate quantification of metals with fluorescent approaches is challenging due to the difficulty in establishing a standard calibration curve in living cells. Herein, a fluorescence imaging protocol is developed for imaging intracellular Cu2+ and its correlation with the cellular uptake of copper. The total amount of intracellular Cu is detected by inductively coupled plasma mass spectrometry (ICP-MS) in parallel. Fluorescence imaging of Cu2+ is accomplished with Rhodamine B derivative modified carbon dots (CDs-Rbh) based on fluorescence resonance energy transfer (FRET) from CDs to rhodamine. Intracellular Cu2+ is correlated with fluorescence ratio at lambda(em) 500-600 nm (rhodamine) to lambda(em) 425-475 nm (CDs) with excitation at lambda(ex) 405 nm. It is found that Cu2+ is linearly correlated with the total intracellular uptaken copper content, with a linear correlation between the relative fluorescence ratio in fluorescence imaging and intracellular Cu derived from ICP-MS, including both Cu (I) and Cu(II) species. The linear calibration equation is lg(F-2/F-1) = 0.00148 m[Cu]-0.3622. This approach facilitates further investigation and elucidation of copper transition in live cells and the evaluation of their cytotoxicity.
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