Optical dark-field spectroscopy of single plasmonic nanoparticles for molecular biosciences.

An ideal sensor capable of quantifying analytes in minuscule sample volumes represents a significant technological advancement. Plasmonic nanoparticles integrated with optical dark-field spectroscopy have reached this capability, demonstrating versatility and expanding applicability across in vitro and in vivo subjects. This review underscores the applicability of optical dark-field spectroscopy with single plasmonic nanoparticles to elucidate a wide range of biomolecular characteristics, including binding constants, molecular dynamics, distances, and forces, as well as recording cell communication signals. Perspectives highlight the potential for the development of implantable nanosensors for metabolite detection in animal models, illustrating the technique's efficacy without the need for labeling molecules. In summary, this review aims to consolidate knowledge of this adaptable and robust technique for decoding molecular biological phenomena within the nano- and bio-scientific community.

Frequency-Domain Method for Characterization of Upconversion Luminescence Kinetics.

The frequency-domain (FD) method provides an alternative to the commonly used time-domain (TD) approach in characterizing the luminescence kinetics of luminophores, with its own strengths, e.g., the capability to decouple multiple lifetime components with higher reliability and accuracy. While extensively explored for characterizing luminophores with down-shifted emission, this method has not been investigated for studying nonlinear luminescent materials such as lanthanide-doped upconversion nanoparticles (UCNPs), featuring more complicated kinetics. In this work, employing a simplified rate-equation model representing a standard two-photon energy-transfer upconversion process, we thoroughly analyzed the response of the luminescence of UCNPs in the FD method. We found that the FD method can potentially obtain from a single experiment the effective decay rates of three critical energy states of the sensitizer/activator ions involved in the upconversion process. The validity of the FD method is demonstrated by experimental data, agreeing reasonably well with the results obtained by TD methods.

Excitation Pulse Duration Response of Upconversion Nanoparticles and Its Applications.

Lanthanide-doped upconversion nanoparticles (UCNPs) have rich photophysics exhibiting complex luminescence kinetics. In this work, we thoroughly investigated the luminescence response of UCNPs to excitation pulse durations. Analyzing this response opens new opportunities in optical encoding/decoding and the assignment of transitions to emission peaks and provides advantages in applications of UCNPs, e.g., for better optical sectioning and improved luminescence nanothermometry. Our work shows that monitoring the UCNP luminescence response to excitation pulse durations (while keeping the duty cycle constant) by recording the average luminescence intensity using a low-time resolution detector such as a spectrometer offers a powerful approach for significantly extending the utility of UCNPs.

Achieving low-power single-wavelength-pair nanoscopy with NIR-II continuous-wave laser for multi-chromatic probes.

Stimulated emission depletion (STED) microscopy is a powerful diffraction-unlimited technique for fluorescence imaging. Despite its rapid evolution, STED fundamentally suffers from high-intensity light illumination, sophisticated probe-defined laser schemes, and limited photon budget of the probes. Here, we demonstrate a versatile strategy, stimulated-emission induced excitation depletion (STExD), to deplete the emission of multi-chromatic probes using a single pair of low-power, near-infrared (NIR), continuous-wave (CW) lasers with fixed wavelengths. With the effect of cascade amplified depletion in lanthanide upconversion systems, we achieve emission inhibition for a wide range of emitters (e.g., Nd3+, Yb3+, Er3+, Ho3+, Pr3+, Eu3+, Tm3+, Gd3+, and Tb3+) by manipulating their common sensitizer, i.e., Nd3+ ions, using a 1064-nm laser. With NaYF4:Nd nanoparticles, we demonstrate an ultrahigh depletion efficiency of 99.3 ± 0.3% for the 450 nm emission with a low saturation intensity of 23.8 ± 0.4 kW cm-2. We further demonstrate nanoscopic imaging with a series of multi-chromatic nanoprobes with a lateral resolution down to 34 nm, two-color STExD imaging, and subcellular imaging of the immunolabelled actin filaments. The strategy expounded here promotes single wavelength-pair nanoscopy for multi-chromatic probes and for multi-color imaging under low-intensity-level NIR-II CW laser depletion.

Nanojet Trapping of a Single Sub-10 nm Upconverting Nanoparticle in the Full Liquid Water Temperature Range.

Upconverting nanoparticles (UCNPs) have been used as optical probes in a great variety of scenarios ranging from cells to animal models. When optically trapped, a single UCNP can be remotely manipulated making possible, for instance, thermal scanning in the surroundings of a living cell. When conventional optics is used, the stability of an optically trapped UCNP is very limited. Its reduced size leads to optical potentials comparable to thermal energy, and up to now, stable optical trapping of a UCNP has been demonstrated only close to room temperature. This fact limits their use above room temperature, for instance, the use to investigate protein denaturalization that occurs in the 40-50 °C range. In this work, stable optical trapping of a single UCNP in the 20-90 °C range has been demonstrated by using a photonic nanojet. The use of an optically trapped microsphere makes it possible to overcome the diffraction limit producing another optical trap of smaller size and enhanced strength. This simple strategy leads not only to an improvement in the thermal stability of the optical trap but also to an enhancement of the emission intensity generated by the optically trapped UCNP.

Optical Manipulation of Lanthanide-Doped Nanoparticles: How to Overcome Their Limitations.

Since Ashkin's pioneering work, optical tweezers have become an essential tool to immobilize and manipulate microscale and nanoscale objects. The use of optical tweezers is key for a variety of applications, including single-molecule spectroscopy, colloidal dynamics, tailored particle assembly, protein isolation, high-resolution surface studies, controlled investigation of biological processes, and surface-enhanced spectroscopy. In recent years, optical trapping of individual sub-100-nm objects has got the attention of the scientific community. In particular, the three-dimensional manipulation of single lanthanide-doped luminescent nanoparticles is of great interest due to the sensitivity of their luminescent properties to environmental conditions. Nevertheless, it is really challenging to trap and manipulate single lanthanide-doped nanoparticles due to the weak optical forces achieved with conventional optical trapping strategies. This limitation is caused, firstly, by the diffraction limit in the focusing of the trapping light and, secondly, by the Brownian motion of the trapped object. In this work, we summarize recent experimental approaches to increase the optical forces in the manipulation of lanthanide-doped nanoparticles, focusing our attention on their surface modification and providing a critical review of the state of the art and future prospects.

Exploring Single-Nanoparticle Dynamics at High Temperature by Optical Tweezers.

The experimental determination of the velocity of a colloidal nanoparticle (vNP) has recently became a hot topic. The thermal dependence of vNP is still left to be explored although it is a valuable source of information allowing, for instance, the discernment between ballistic and diffusive regimes. Optical tweezers (OTs) constitute a tool especially useful for the experimental determination of vNP although they have only been capable of determining it at room temperature. In this work, we demonstrate that it is possible to determine the temperature dependence of the diffusive velocity of a single colloidal nanoparticle by analyzing the temperature dependence of optical forces. The comparison between experimental results and theoretical predictions allowed us to discover the impact that the anomalous temperature dependence of water properties has on the dynamics of colloidal nanoparticles in this temperature range.

pH dependence of water anomaly temperature investigated by Eu(III) cryptate luminescence.

Although water has been extensively studied, not all of its unique properties have been fully understood. There is still controversy about the temperature at which hydrogen bonds are broken or weakened, producing the anomalous temperature dependence of many water properties. Different temperatures between 23 and 48 °C have been reported, but no study has scrutinized the reasons for this discrepancy. We suggest the determining role of pH in the alteration of the water anomaly temperature. We employed a luminescent europium trisbipyridine cryptate, which is highly sensitive to changes in the arrangement of water molecules and whose luminescence intensity and lifetime are not significantly influenced by variations over a broad pH range. Our results revealed an increase of the crossover temperature from circa 35 °C at pH 3.5 to circa 45 °C at pH 7 to 9, which explains the discrepancies of previous studies. The pH dependence of water anomaly temperature is an important property for a better understanding of water and water-based systems and applications.

Reliability of rare-earth-doped infrared luminescent nanothermometers.

The use of infrared-emitting rare-earth-doped luminescent nanoparticles as nanothermometers has attracted great attention during the last few years. The scientific community has identified rare-earth-doped luminescent nanoparticles as one of the most sensitive and versatile systems for contactless local temperature sensing in a great variety of fields, but especially in nanomedicine. Researchers are nowadays focused on the design and development of multifunctional nanothermometers with new spectral operation ranges, outstanding brightness, and enhanced sensitivities. However, no attention has been paid to the assessment of the actual reliability of the measurements provided by rare-earth-doped luminescent nanothermometers. In fact, it is assumed that they are ideal temperature sensors. Nevertheless, this is far from being true. In this work we demonstrate that the emission spectra of rare-earth-doped nanothermometers can be affected by numerous environmental and experimental factors. These include the numerical aperture of the optical elements used for their optical excitation and luminescence collection, the local concentration of nanothermometers, optical length variations, self-absorption of the luminescence by the nanothermometers themselves, and solvent optical absorption. This work concludes that rare-earth-doped luminescent nanothermometers are not as reliable as thought and, consequently, special care has to be taken when extracting temperature estimations from the variation of their emission spectra.

Core-shell rare-earth-doped nanostructures in biomedicine.

The current status of the use of core-shell rare-earth-doped nanoparticles in biomedical applications is reviewed in detail. The different core-shell rare-earth-doped nanoparticles developed so far are described and the most relevant examples of their application in imaging, sensing, and therapy are summarized. In addition, the advantages and disadvantages they present are discussed. Finally, a critical opinion of their potential application in real life biomedicine is given.

Upconverting nanocomposites with combined photothermal and photodynamic effects.

Lanthanide-doped upconverting nanoparticles (UCNPs) have been studied for diverse biomedical applications due to their inherent ability to convert near-infrared (NIR) excitation light to higher energies (spanning the ultraviolet, visible, and NIR regions). To explore additional functionalities, rational combination with other optically active nanostructures may lead to the development of new multimodal nanoplatforms with theranostic (therapy and diagnostic) capabilities. Here, we develop a nanocomposite consisting of NaGdF4:Er3+, Yb3+ UCNPs, mesoporous silica (SiO2), gold nanorods (GNRs) and a photosensitizer, with integrated functionalities including luminescence imaging, photothermal generation, nanothermometry and photodynamic effects. Under 980 nm irradiation, GNRs and UCNPs are simultaneously excited due to the overlap between the surface plasmon resonance of the GNRs and the absorption of the UCNPs leading to plasmonic enhancement of the upconverted luminescence, while concomitantly creating a temperature gradient. The temperature increase can be determined from the intensity ratio of the upconverted green emission of the UCNPs. Finally, a photosensitizer, zinc phthalocyanine, was loaded into the mesoporous SiO2. Upon laser irradiation, the upconverted visible light subsequently activates the photosensitizer to release reactive oxygen species. The multifunctional GNR@SiO2@UCNPs nanocomposites showed strong luminescence signal when incubated in HeLa cervical cancer cells, making them ideal bioprobes for future theranostic applications.

Unveiling Molecular Changes in Water by Small Luminescent Nanoparticles.

Nowadays a large variety of applications are based on solid nanoparticles dispersed in liquids-so called nanofluids. The interaction between the fluid and the nanoparticles plays a decisive role in the physical properties of the nanofluid. A novel approach based on the nonradiative energy transfer between two small luminescent nanocrystals (GdVO4 :Nd3+ and GdVO4 :Yb3+ ) dispersed in water is used in this work to investigate how temperature affects both the processes of interaction between nanoparticles and the effect of the fluid on the nanoparticles. From a systematic analysis of the effect of temperature on the GdVO4 :Nd3+ → GdVO4 :Yb3+ interparticle energy transfer, it can be concluded that a dramatic increase in the energy transfer efficiency occurs for temperatures above 45 °C. This change is properly explained by taking into account a crossover existing in diverse water properties that occurs at about this temperature. The obtained results allow elucidation on the molecular arrangement of water molecules below and above this crossover temperature. In addition, it is observed that an energy transfer process is produced as a result of interparticle collisions that induce irreversible ion exchange between the interacting nanoparticles.

Liquid whispering-gallery-mode resonator as a humidity sensor.

We experimentally demonstrate the high sensitivity of a novel liquid state, whispering-gallery-mode optical resonator to humidity changes. The optical resonator used consists of a droplet made of glycerol, a transparent liquid that enables high optical quality factor, doped with fluorescent material. As glycerol is highly hygroscopic, the refractive index and radius of the droplet change with ambient humidity. This produces a shift on the whispering gallery mode's wavelengths, which modulates the emission of the fluorescent material. This device shows an unpreceded sensitivity of 10-3 per relative humidity percent.

Determining the 3D orientation of optically trapped upconverting nanorods by in situ single-particle polarized spectroscopy.

An approach to unequivocally determine the three-dimensional orientation of optically manipulated NaYF4:Er(3+),Yb(3+) upconverting nanorods (UCNRs) is demonstrated. Long-term immobilization of individual UCNRs inside single and multiple resonant optical traps allow for stable single UCNR spectroscopy studies. Based on the strong polarization dependent upconverted luminescence of UCNRs it is possible to unequivocally determine, in real time, their three-dimensional orientation when optically trapped. In single-beam traps, polarized single particle spectroscopy has concluded that UCNRs orientate parallel to the propagation axis of the trapping beam. On the other hand, when multiple-beam optical tweezers are used, single particle polarization spectroscopy demonstrated how full spatial control over UCNR orientation can be achieved by changing the trap-to-trap distance as well as the relative orientation between optical traps. All these results show the possibility of real time three-dimensional manipulation and tracking of anisotropic nanoparticles with wide potential application in modern nanobiophotonics.