Key Laboratory of Optic-electric Sensing and Analytical Chemistry for Life Science, MOE; State Key Laboratory Base for Eco-chemical Engineering; Shandong Key Laboratory of Biochemical Analysis; Key Laboratory of Analytical Chemistry for Life Science in Universities of Shandong; College of Chemistry and Molecular Engineering . Qingdao University of Science and Technology , Qingdao 266042 , PR China.
ACS Sens. 2019 Apr 26;4(4):924-930. doi: 10.1021/acssensors.9b00031. Epub 2019 Apr 10.
Imaging and detecting microRNAs (miRNAs) is of central importance in tumor cell analysis. It stays challenging to establish simple, accurate, and sensitive analytical assays for imaging and detection of miRNA in a single living cell, because of intracellular complex environment and miRNA sequence similarity. Herein, we designed a dual-signal twinkling probe (DSTP) with triplex-stem structure which employed a fluorescence-SERS signal reciprocal switch. The spatiotemporal dynamics of the miRNA molecular and intracellular uptake of the probe are monitored by fluorescence-SERS signal switch of the DSTP. Meanwhile, using the surface-enhanced Raman scattering (SERS) signals of DSTP, the measure of absolute value coupling of reciprocal signals is first used to real-time detection of miRNA. Through simultaneous enhancing the target response signal value and reducing blank value, this work deducted the background effect, and showed high sensitivity and reproducibility. Moreover, the probe shows excellent reversibility and specificity in real quantitative detection of intracellular miRNA. miR-203 was successfully monitored in MCF-7, in accord with the results in vitro as well as in cell lysates. We anticipate that this new dual-signal twinkling and dual-spectrum switch method will be generally useful to image and detect various types of biomolecules in single living cell.
在肿瘤细胞分析中,对 microRNAs(miRNAs)进行成像和检测至关重要。由于细胞内复杂的环境和 miRNA 序列的相似性,建立简单、准确和灵敏的分析方法来对单个活细胞中的 miRNA 进行成像和检测仍然具有挑战性。在此,我们设计了一种具有三链茎结构的双信号闪烁探针(DSTP),它利用荧光-SERS 信号相互切换。通过 DSTP 的荧光-SERS 信号开关来监测 miRNA 分子的时空动力学和探针的细胞内摄取。同时,利用 DSTP 的表面增强拉曼散射(SERS)信号,首次使用相互信号的绝对值耦合的测量来实时检测 miRNA。通过同时增强目标响应信号值并降低空白值,本工作消除了背景效应,表现出了高灵敏度和重现性。此外,该探针在细胞内 miRNA 的实时定量检测中表现出优异的可逆性和特异性。miR-203 在 MCF-7 中成功监测,与体外以及细胞裂解物中的结果一致。我们预计这种新的双信号闪烁和双光谱切换方法将广泛适用于单个活细胞中各种类型的生物分子的成像和检测。