The "fixed" or "flexible" design of plasmonic hotspots is a frontier area of research in the field of surface-enhanced Raman scattering (SERS). Most reported SERS hotspots have been shown to exist in zero-dimensional point-like, one-dimensional linear, or two-dimensional planar geometries. Here, we demonstrate a novel three-dimensional (3D) hotspot matrix that can hold hotspots between every two adjacent particles in 3D space, simply achieved by evaporating a droplet of citrate-Ag sols on a fluorosilylated silicon wafer. In situ synchrotron-radiation small-angle X-ray scattering (SR-SAXS), combined with dark-field microscopy and in situ micro-UV, was employed to explore the evolution of the 3D geometry and plasmonic properties of Ag nanoparticles in a single droplet. In such a droplet, there is a distinct 3D geometry with minimal polydispersity of particle size and maximal uniformity of interparticle distance, significantly different from the dry state. According to theoretical simulations, the liquid adhesive force promotes a closely packed assembly of particles, and the interparticle distance is not fixed but can be balanced in a small range by the interplay of the van der Waals attraction and electrostatic repulsion experienced by a particle. The "trapping well" for immobilizing particles in 3D space can result in a large number of hotspots in a 3D geometry. Both theoretical and experimental results demonstrate that the 3D hotspots are predictable and time-ordered in the absence of any sample manipulation. Use of the matrix not only produces giant Raman enhancement at least 2 orders of magnitude larger than that of dried substrates, but also provides the structural basis for trapping molecules. Even a single molecule of resonant dye can generate a large SERS signal. With a portable Raman spectrometer, the detection capability is also greatly improved for various analytes with different natures, including pesticides and drugs. This 3D hotspot matrix overcomes the long-standing limitations of SERS for the ultrasensitive characterization of various substrates and analytes and promises to transform SERS into a practical analytical technique.

As acute myocardial infarction (AMI) has now become a severe death threat to humans and may abruptly occur at home or outdoors where sophisticated equipment is not available, it is of great importance to develop facile methodologies for the point-of-care (POC) diagnosis of AMI. Toward this goal, here we build a sensing platform for chemiluminescence (CL) microRNA (miRNA) imaging with a smartphone as the portable detector, and for the first time we achieve visualization of AMI-related miRNAs in real patients' serum. We first construct a spherical nucleic acid enzyme (termed SNAzyme) derived from a dense layer of G-quadruplex (G4) DNAzyme formed on the gold nanoparticle core, which displays ∼100-fold and higher catalytic activity and improved resistance to nuclease degradation in a real blood sample as compared to those of the G4 DNAzyme itself. These unique features endow the SNAzyme-boosted CL platform with superior imaging performance for analyzing an AMI-related miRNA, miRNA-133a. This miRNA is employed to trigger the target-catalyzed hairpin assembly to produce a sticky dsDNA linker that captures the SNAzyme nanolabel onto the substrate. In this way, miRNA-133a is successfully detected, with a limit of detection of 0.3 pM (S/N = 3) and a high selectivity over other miRNA analogs in patients' blood. Given its unique features in physiological environments, our SNAzyme-boosted imaging platform holds great promise for use in the POC diagnosis of AMI.

SERS hot spots with outstanding enhancement ability can spontaneously form in a reversible and reproducible way by the self-approach of flexible silver nanorods driven by the capillary force of solvent evaporation, and at the same time the target analytes can be trapped in the top-closed silver nanorods in the solvent evaporation process. The Raman intensity of the top-closed vs. top-opened nanorod arrays was a factor of 100-1000 higher for SERS reporters such as p-aminothiophenol or crystal violet.Furthermore, triplicate measurements on long nanorod arrays at the same position show a variation of the Raman intensity <10%, demonstrating a good reproducibility of the enhancement. Moreover, we found that the self-approach is highly dependent on the nanorod length and the molecules with different adsorptivity have different SERS performance in the solvent evaporation process. This solvent evaporation-controlled self-approach is an extremely simple and efficient strategy for the spontaneous formation of Raman hot spots with outstanding enhancement ability. These characteristics promise a generic platform for molecule trapping and SERS sensing with high sensitivity and reproducibility, which can help to transform SERS into a practical analytical technique.