Fluorescence and SERS Imaging for the Simultaneous Absolute Quantification of Multiple miRNAs in Living Cells
Fluorescence and SERS Imaging for the Simultaneous Absolute Quantification of Multiple miRNAs in Living Cells
复制标题
荧光和 SERS 成像可同时绝对定量活细胞中的多种 miRNA
DOI：
10.1021/acs.analchem.7b00697
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发表时间：
2017-05-02
影响因子：
7.4
通讯作者：
Tang, Bo
中科院分区：
文献类型：
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作者：
Ye, Sujuan;Li, Xiaoxiao;Tang, Bo
多个细胞内微小RNA(MiRNAs)的同时成像和定量对于癌症的早期诊断尤为必要。然而，同时直接成像和绝对定量多个细胞内RNA仍然是一个巨大的挑战，特别是对于在活细胞中表达水平显著不同的miRNAs来说。我们利用荧光-拉曼信号开关设计了双信号可开关(DSS)纳米探针。该探针的细胞内摄取和动态行为通过其荧光信号进行监测。同时，通过测量表面增强拉曼光谱(SERS)比率，实时定量检测多个miRNAs成为可能。此外，信号1：N比率放大模式只响应低丰度的miRNA(不对称信号放大模式)，用于同时可视化和定量检测活细胞中显著不同水平的miRNAs。在活的MCF-7细胞中成功地检测到了miR-21和miR-203，这与同一批细胞裂解物的体外结果一致。已报道的双光谱成像方法有望为各种生物分子的细胞内成像和检测提供一种新的策略。
The simultaneous imaging and quantification of multiple intracellular microRNAs (miRNAs) are particularly desirable for the early diagnosis of cancers. However, simultaneous direct imaging with absolute quantification of multiple intracellular RNAs remains a great challenge, particularly for miRNAs, which have significantly different expression levels in living cells. We designed dual-signal switchable (DSS) nanoprobes using the fluorescence-Raman signal switch. The intracellular uptake and dynamic behaviors of the probe are monitored by its fluorescence signal. Meanwhile, real-time quantitative detection of multiple miRNAs is made possible by measurements of the surface-enhanced Raman spectroscopy (SERS) ratios. Moreover, the signal 1:n ratio amplification mode only responds to low abundance miRNA (asymmetric signal amplification mode) for simultaneous visualization and quantitative detection of significantly different levels of miRNAs in living cells. miR-21 and miR-203 were successfully detected in living MCF-7 cells, in agreement with in vitro results from the same batch of cell lysates. The reported dual-spectrum imaging method promises to offer a new strategy for the intracellular imaging and detection of various types of biomolecules.