Multiplexed intracellular detection based on dual-excitation/dual-emission upconversion nanoprobes
upconversion nanoparticles, energy transfer, dual-excitation, dye sensitization, ratiometric probe, multiplexed detection
Multiplexed intracellular detection is desirable in biomedical sciences for its higher efficiency and accuracy compared to the singleanalyte detection. However, it is very challenging to construct nanoprobes that possess multiple fluorescent signals to recognize the different intracellular species synchronously. Herein, we proposed a novel dual-excitation/dual-emission upconversion strategy for multiplexed detection through the design of upconversion nanoparticles (UCNP) loaded with two dyes for sensitization and quenching of the upconversion luminescence (UCL), respectively. Based on the two independent energy transfer processes of near-infrared (NIR) dye IR845 to UCNP and UCNP to visible dye PAPS-Zn, ClO− and Zn2+ were simultaneously detected with a limit of detection (LOD) of 41.4 and 10.5 nM, respectively. By utilizing a purpose-built 830/980 nm dual-laser confocal microscope, both intrinsic and exogenous ClO− and Zn2+ in live MCF-7 cells have been accurately quantified. Such dual-excitation/dual-emission ratiometric UCL detection mode enables not only monitoring multiple intracellular analytes but also eliminating the detection deviation caused by inhomogeneous probe distribution in cells. Through modulation of NIR dye and visible dye with other reactive groups, the nanoprobes can be extended to analyze various intracellular species, which provides a promising tool to study the biological activities in live cells and diagnose diseases.
Tsinghua University Press
Jianxi Ke,Shan Lu,Zhuo Li,Xiaoying Shang,Xingjun Li,Renfu Li,Datao Tu,Zhuo Chen,Xueyuan Chen, Multiplexed intracellular detection based on dual-excitation/dual-emission upconversion nanoprobes. NanoRes.2020, 13(7): 1955–1961