Key Laboratory of Optic-electric Sensing and Analytical Chemistry for Life Science, MOE; Shandong Key Laboratory of Biochemical Analysis; College of Chemistry and Molecular Engineering. Qingdao University of Science and Technology, Qingdao 266042, P. R. China.
College of Chemistry, Chemical Engineering and Materials Science, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Shandong Normal University, Jinan 250014, P. R. China.
Anal Chem. 2021 Jan 26;93(3):1466-1471. doi: 10.1021/acs.analchem.0c03491. Epub 2020 Dec 21.
The expression of microRNAs (miRNAs) is critical in gene regulation and has been counted into disease diagnosis marks. Precise imaging and quantification of miRNAs could afford the important information for clinical diagnosis. Here, two smart binary star ratio (BSR) probes were designed and constructed, and miRNA triggered the connection of the binary star probes and the reciprocal changes of dual signals in living cells. This multifunctional probe integrates fluorescence and surface enhanced Raman scattering (SERS) imaging, with enzyme-free numerator signal amplification for dual-mode imaging and dual-signal quantitative analysis of miRNA. First, compared with the single-mode ratio imaging method, using fluorescence-SERS complementary ratio imaging, this probe enables more accurate imaging contrast for direct visualization signal changes in living cells. Multiscale information about the dynamic behavior of miRNA and the probe is acquired. Next, via SERS reverse signal ratio response and a novel enzyme-free numerator signal amplification, the amplified signal and reduced black value were achieved in the quantification of miRNA. More importantly, BSR probes showed good stability in cells and were successfully used for accurate tracing and quantification of miR-203 from MCF-7 cells. Therefore, the reported BSR probe is a potential tool for the reliable monitoring of biomolecule dynamics in living cells.
miRNAs 的表达在基因调控中至关重要，并被认为是疾病诊断的标志物。miRNAs 的精确成像和定量可以为临床诊断提供重要信息。在这里，我们设计并构建了两种智能双星比率（BSR）探针，miRNA 触发了双星探针的连接以及活细胞中双信号的相互变化。这种多功能探针集成了荧光和表面增强拉曼散射（SERS）成像，具有无酶分子信号扩增功能，用于双模式成像和 miRNA 的双信号定量分析。首先，与单模比率成像方法相比，使用荧光-SERS 互补比率成像，该探针能够更准确地对活细胞中直接可视化信号变化进行成像对比。获得了有关 miRNA 和探针的动态行为的多尺度信息。接下来，通过 SERS 反向信号比率响应和新颖的无酶分子信号扩增，实现了 miRNA 定量分析中的信号放大和黑值降低。更重要的是，BSR 探针在细胞中表现出良好的稳定性，并成功用于 MCF-7 细胞中 miR-203 的精确跟踪和定量。因此，所报道的 BSR 探针是一种用于可靠监测活细胞中生物分子动力学的潜在工具。
