Intracellular Zinc Quantification by Fluorescence Imaging with a FRET System

Analytical Chemistry
Yang ShuJian-Hua Wang

Abstract

Fluorescence imaging of cellular metals is widely reported. However, the quantification of intracellular metals with fluorescence imaging is so far not feasible and highly challenging. In this work, a ratiometric probe with two fluorescently labeled complementary DNA strains is designed for intracellular zinc quantification via fluorescence imaging, based on fluorescence resonance energy transfer (FRET) from carbon dots (CDs) to fluorescein (FAM). The donor CDs are modified with a Zn2+ aptamer, whereas the receptor FAM is conjugated with the complementary DNA sequence to ensure selectivity. MCF-7 cells are cultured sequentially with Zn2+ (20, 40, 55, 70, 85, and 100 μmol L-1) and CDs-FAM (100 μg mL-1), which is used for fluorescence imaging (at λex = 405 nm and λem = 440-490 nm for CDs, λem = 500-550 nm for FAM) to provide a relative fluorescence ratio (( F - F0)/ F0, F = ICDs/ IFAM), followed by quantifying intracellular zinc with ICPMS. A linear correlation is achieved between the relative fluorescence ratio in fluorescence images and the intracellular zinc content derived by ICPMS, which facilitates intracellular zinc quantification via fluorescence imaging. It is especially useful for real-time tracing of intracellular zinc...Continue Reading

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Citations

Feb 11, 2020·Wiley Interdisciplinary Reviews. Nanomedicine and Nanobiotechnology·Haydar AliNikhil R Jana
Feb 6, 2020·Topics in Current Chemistry·Yuan ZhaoChuanlai Xu
Oct 16, 2021·ACS Applied Materials & Interfaces·Ya-Ting GaoMahmoud Elsayed Hafez
Dec 17, 2021·Chemical Communications : Chem Comm·Xuanhao ZhangXiliang Luo

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