Implantable Sensors Based on Gold Nanoparticles for Continuous Long-Term Concentration Monitoring in the Body.

Implantable sensors continuously transmit information on vital values or biomarker concentrations in bodily fluids, enabling physicians to survey disease progression and monitor therapeutic success. However, currently available technologies still face difficulties with long-term operation and transferability to different analytes. We show the potential of a generalizable platform based on gold nanoparticles embedded in a hydrogel for long-term implanted biosensing. Using optical imaging and an intelligent sensor/reference-design, we assess the tissue concentration of kanamycin in anesthetized rats by interrogating our implanted sensor noninvasively through the skin. Combining a tissue-integrating matrix, robust aptamer receptors, and photostable gold nanoparticles, our technology has strong potential to extend the lifetime of implanted sensors. Because of the easy adaptability of gold nanoparticles toward different analytes, our concept will find versatile applications in personalized medicine or pharmaceutical development.

Single Out-of-Resonance Dielectric Nanoparticles as Molecular Sensors.

Light scattering from single nanoparticles and nanostructures is a commonly used readout method for nanosensors. Increasing the spectral sensitivity of resonant nanosensors to changes in their local surrounding has been the focus of many studies. Switching from spectral to intensity monitoring allows one to investigate nonresonant or out-of-resonance dielectric nanoparticles. Here, we systematically compared such dielectric silica nanoparticles with plasmonic gold nanorods by deriving analytical expressions and by performing experiments. The experiments show a similar sensitivity for the detection of an adsorbate layer for both particle types, which is in good agreement with theory. The flat spectral response of dielectric silica nanoparticles simplifies the choice of illumination wavelength. Furthermore, such dielectric nanoparticles can be made from many oxides, polymers, and even biological assemblies, broadening the choice of materials for the nanosensor.

Intensity-Based Single Particle Plasmon Sensing.

Plasmon sensors respond to local changes of their surrounding environment with a shift in their resonance wavelength. This response is usually detected by measuring light scattering spectra to determine the resonance wavelength. However, single wavelength detection has become increasingly important because it simplifies the setup, increases speed, and improves statistics. Therefore, we investigated theoretically how the sensitivity toward such single wavelength scattering intensity changes depend on the material and shape of the plasmonic sensor. Surprisingly, simple equations describe this intensity sensitivity very accurately and allow us to distinguish the various contributions: Rayleigh scattering, dielectric contrast, plasmon shift, and frequency-dependent plasmon bulk damping. We find very good agreement of theoretical predictions and experimental data obtained by single particle spectroscopy.

Plasmonic Nanosensors for the Label-Free Imaging of Dynamic Protein Patterns.

We introduce a new approach to monitor the dynamics and spatial patterns of biological molecular assemblies. Our molecular imaging method relies on plasmonic gold nanoparticles as point-like detectors and requires no labeling of the molecules. We show spatial resolution of up to 5 µm and 30 ms temporal resolution, which is comparable to wide-field fluorescence microscopy, while requiring only readily available gold nanoparticles and a dark-field optical microscope. We demonstrate the method on MinDE proteins attaching to and detaching from lipid membranes of different composition for 24 h. We foresee our new imaging method as an indispensable tool in advanced molecular biology and biophysics laboratories around the world.

Plasmonic Nanosensors Reveal a Height Dependence of MinDE Protein Oscillations on Membrane Features.

Single-particle plasmon spectroscopy has become a standard technique to detect and quantify the presence of unlabeled macromolecules. Here, we extend this method to determine their exact distance from the plasmon sensors with sub-nanometer resolution by systematically varying the sensing range into the surrounding by adjusting the size of the plasmonic nanoparticles. We improved current single-particle plasmon spectroscopy to record continuously for hours the scattering spectra of thousands of nanoparticles of different sizes simultaneously with 1.8 s time resolution. We apply this technique to study the interaction dynamics of bacterial Min proteins with supported lipid membranes of different composition. Our experiments reveal a surprisingly flexible operating mode of the Min proteins: In the presence of cardiolipin and membrane curvature induced by nanoparticles, the protein oscillation occurs on top of a stationary MinD patch. Our results reveal the need to consider membrane composition and local curvature as important parameters to quantitatively understand the Min protein system and could be extrapolated to other macromolecular systems. Our label-free method is generally easily implementable and well suited to measure distances of interacting biological macromolecules.

Conformational Dynamics of a Single Protein Monitored for 24 h at Video Rate.

We use plasmon rulers to follow the conformational dynamics of a single protein for up to 24 h at a video rate. The plasmon ruler consists of two gold nanospheres connected by a single protein linker. In our experiment, we follow the dynamics of the molecular chaperone heat shock protein 90 (Hsp90), which is known to show "open" and "closed" conformations. Our measurements confirm the previously known conformational dynamics with transition times in the second to minute time scale and reveals new dynamics on the time scale of minutes to hours. Plasmon rulers thus extend the observation bandwidth 3-4 orders of magnitude with respect to single-molecule fluorescence resonance energy transfer and enable the study of molecular dynamics with unprecedented precision.

Chemical Interface Damping Depends on Electrons Reaching the Surface.

Metallic nanoparticles show extraordinary strong light absorption near their plasmon resonance, orders of magnitude larger compared to nonmetallic nanoparticles. This "antenna" effect has recently been exploited to transfer electrons into empty states of an attached material, for example to create electric currents in photovoltaic devices or to induce chemical reactions. It is generally assumed that plasmons decay into hot electrons, which then transfer to the attached material. Ultrafast electron-electron scattering reduces the lifetime of hot electrons drastically in metals and therefore strongly limits the efficiency of plasmon induced hot electron transfer. However, recent work has revived the concept of plasmons decaying directly into an interfacial charge transfer state, thus avoiding the intermediate creation of hot electrons. This direct decay mechanism has mostly been neglected, and has been termed chemical interface damping (CID). CID manifests itself as an additional damping contribution to the homogeneous plasmon line width. In this study, we investigate the size dependence of CID by following the plasmon line width of gold nanorods during the adsorption process of thiols on the gold surface with single particle spectroscopy. We show that CID scales inversely with the effective path length of electrons, i.e., the average distance of electrons to the surface. Moreover, we compare the contribution of CID to other competing plasmon decay channels and predict that CID becomes the dominating plasmon energy decay mechanism for very small gold nanorods.

Detection and imaging of quorum sensing in Pseudomonas aeruginosa biofilm communities by surface-enhanced resonance Raman scattering.

Most bacteria in nature exist as biofilms, which support intercellular signalling processes such as quorum sensing (QS), a cell-to-cell communication mechanism that allows bacteria to monitor and respond to cell density and changes in the environment. As QS and biofilms are involved in the ability of bacteria to cause disease, there is a need for the development of methods for the non-invasive analysis of QS in natural bacterial populations. Here, by using surface-enhanced resonance Raman scattering spectroscopy, we report rationally designed nanostructured plasmonic substrates for the in situ, label-free detection of a QS signalling metabolite in growing Pseudomonas aeruginosa biofilms and microcolonies. The in situ, non-invasive plasmonic imaging of QS in biofilms provides a powerful analytical approach for studying intercellular communication on the basis of secreted molecules as signals.

The role of halide ions in the anisotropic growth of gold nanoparticles: a microscopic, atomistic perspective.

We provide a microscopic view of the role of halides in controlling the anisotropic growth of gold nanorods through a combined computational and experimental study. Atomistic molecular dynamics simulations unveil that Br(-) adsorption is not only responsible for surface passivation, but also acts as the driving force for CTAB micelle adsorption and stabilization on the gold surface in a facet-dependent way. The partial replacement of Br(-) by Cl(-) decreases the difference between facets and the surfactant density. Finally, in the CTAC solution, no halides or micellar structures protect the gold surface and further gold reduction should be uniformly possible. Experimentally observed nanoparticle's growth in different CTAB/CTAC mixtures is more uniform and faster as the amount of Cl(-) increases, confirming the picture from the simulations. In addition, the surfactant layer thickness measured on nanorods exposed to CTAB and CTAC quantitatively agrees with the simulation results.