Fluorescence and SERS Imaging for the Simultaneous Absolute Quantification of Multiple miRNAs in Living Cells

Abstract

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.

Accurate cancer cell identification and microRNA silencing induced therapy using tailored DNA tetrahedron nanostructures

“…With the development of nanomaterials, a variety of nanoprobes have been reported for the detection of multiple miRNAs. [9][10][11][12] For example, the Tang group developed multicolor uorescent nanoprobes based on gold nanoparticles (GNPs) for evaluating cellular migration and invasion by simultaneously imaging miRNA-221, PTEN mRNA and MMP9 in living cells. 13 The Zhu group assembled multicomponent nucleic acid enzymes onto the surface of mesoporous silica-coated gold nanorods as multifunctional nanodevices for intracellular miRNA-21 and miRNA-145 in situ imaging.…”

Sensing of Vimentin mRNA in 2D and 3D Models of Wounded Skin Using DNA‐Coated Gold Nanoparticles

“…[17] Pioneering work by Mirkin and co-workers as well as Tang and co-workers recently introduced a new type of fluorescent, oligonucleotide-based imaging probe immobilized on a gold nanoparticle (AuNP), the so-called nanoflares, for intracellular mRNA detection and gene regulation. [18][19][20][21][22][23][24] Recently, our group developed this strategy further to transport drugs into cells and release them only in the presence of a specific mRNA signature. [25] This approach allowed us to selectively sense model tumor cells and trigger the release of drug payloads leading to efficient cell death, while other cell types remained unaffected.…”

A pH ratiometrically responsive surface enhanced resonance Raman scattering probe for tumor acidic margin delineation and image-guided surgery

“…21 The second is the incorporation of two or more types of Raman reporters into a particle, in which one inert reporter serves as an internal reference and the others are used for analyte determination. 22 The third is conjugation of a single reporter molecule being functionalized as both an analyte sensor and a self-built-in calibrator. As far as we are aware, AuS-IR7p is the rst Raman probe independently determining physiological pH values with a signal reporter molecule.…”