Integration of a superconducting nanowire single-photon detector into a confocal microscope for time-resolved photoluminescence (TRPL)-mapping: Sensitivity and time resolution.

This report highlights the combination of the MicroTime 100 upright confocal fluorescence lifetime microscope with a Single Quantum Eos Superconducting Nanowire Single-Photon Detector (SNSPD) system as a powerful tool for photophysical research and applications. We focus on an application in materials science, photoluminescence imaging, and lifetime characterization of Cu(InGa)Se2 (CIGS) devices intended for solar cells. We demonstrate improved sensitivity, signal-to-noise ratio, and time-resolution in combination with confocal spatial resolution in the near-infrared (NIR) range, specifically in the 1000-1300 nm range. The MicroTime 100-Single Quantum Eos system shows two orders of magnitude higher signal-to-noise ratio for CIGS devices' photoluminescence imaging compared to a standard NIR-photomultiplier tube (NIR-PMT) and a three-fold improvement in time resolution, which is now limited by the laser pulse width. Our results demonstrate the advantages in terms of image quality and time resolution of SNSPDs technology for imaging in materials science.

Effective Electron Temperature Measurement Using Time-Resolved Anti-Stokes Photoluminescence.

Anti-Stokes photoluminescence of metal nanoparticles, in which emitted photons have a higher energy than the incident photons, is an indicator of the temperature prevalent within a nanoparticle. Previous work has shown how to extract the temperature from a gold nanoparticle under continuous-wave monochromatic illumination. We extend the technique to pulsed illumination and introduce pump-probe anti-Stokes spectroscopy. This new technique enables us not only to measure an effective electron temperature in a gold nanoparticle (∼103 K under our conditions), but also to measure ultrafast dynamics of a pulse-excited electron population, through its effect on the photoluminescence, with subpicosecond time resolution. We measure the heating and cooling, all within picoseconds, of the electrons and find that, with our subpicosecond pulses, the highest apparent temperature is reached 0.6 ps before the maximum change in magnitude of the extinction signal.

Circular Dichroism Measurement of Single Metal Nanoparticles Using Photothermal Imaging.

Circular dichroism (CD) spectroscopy is a powerful optical technique for the study of chiral materials and molecules. It gives access to an enantioselective signal based on the differential absorption of right and left circularly polarized light, usually obtained through polarization analysis of the light transmitted through a sample of interest. CD is routinely used to determine the secondary structure of proteins and their conformational state. However, CD signals are weak, limiting the use of this powerful technique to ensembles of many molecules. Here, we experimentally realize the concept of photothermal circular dichroism, a technique that combines the enantioselective signal from circular dichroism with the high sensitivity of photothermal microscopy, achieving a superior signal-to-noise ratio to detect chiral nano-objects. As a proof of principle, we studied the chiral response of single plasmonic nanostructures with CD in the visible range, demonstrating a signal-to-noise ratio better than 40 with only 30 ms integration time for these nanostructures. The high signal-to-noise ratio allows us to quantify the CD signal for individual nanoparticles. We show that we can distinguish relative absorption differences for right circularly and left circularly polarized light as small as gmin = 4 × 10-3 for a 30 ms integration time with our current experimental settings. The enhanced sensitivity of our technique extends CD studies to individual nano-objects and opens CD spectroscopy to numbers of molecules much lower than those in conventional experiments.

Plasmonic Enhancement of Two-Photon-Excited Luminescence of Single Quantum Dots by Individual Gold Nanorods.

Plasmonic enhancement of two-photon-excited fluorescence is not only of fundamental interest but also appealing for many bioimaging and photonic applications. The high peak intensity required for two-photon excitation may cause shape changes in plasmonic nanostructures, as well as transient plasmon broadening. Yet, in this work, we report on strong enhancement of the two-photon-excited photoluminescence of single colloidal quantum dots close to isolated chemically synthesized gold nanorods. Upon resonant excitation of the localized surface plasmon resonance, a gold nanorod can enhance the photoluminescence of a single quantum dot more than 10 000-fold. This strong enhancement arises from the combined effect of local field amplification and the competition between radiative and nonradiative decay rate enhancements, as is confirmed by time-resolved fluorescence measurements and numerical simulations.

Single-molecule fluorescence enhancement of a near-infrared dye by gold nanorods using DNA transient binding.

Fluorescence enhancement by plasmonic nanostructures enables the optical detection of single molecules with weak fluorescence, extending the scope of molecular fluorescence imaging to new materials and systems. In this work, we study single-molecule fluorescence enhancement by individual gold nanorods exploiting a DNA-based transient binding technique. Single molecules are attached to short DNA oligomers that can reversibly hybridize to their complementary docking DNA strands immobilized on the surface of gold nanorods or the glass substrate next to gold nanorods. This method continuously refreshes the single molecule in the near field of the gold nanorod, and enables a study of fluorescence enhancement at a well-defined position, with long dwell time and without limitation by photobleaching. Docking strands attached to the glass substrate are found to be more photo-stable. We find over 3000-fold fluorescence enhancement of single molecules of IRDye800CW, a near-infrared dye with a low quantum yield of 7%. This strong enhancement, consistent with numerical simulations, arises from the combined effect of local field enhancement and the competition between radiative and nonradiative decay rate enhancements.

Quantifying fluorescence enhancement for slowly diffusing single molecules in plasmonic near fields.

Gold nanorods are extensively used for single-molecule fluorescence enhancement as they are easy to synthesize, bio-compatible, and provide high light confinement at their nanometer-sized tips. The current way to estimate fluorescence enhancement relies on binned time traces or on fluorescence correlation spectroscopy. We report on novel ways to extract the enhancement factor in a single-molecule enhancement experiment, avoiding the arbitrary selection of one or a few high-intensity burst(s). These new estimates for the enhancement factor make use of the whole distribution of intensity bursts or of the interphoton delay distribution, which avoids the arbitrary binning of the fluorescence intensity time traces. We present experimental results on the bi-dimensional case, experimentally achieved using a lipid bilayer to support the diffusion of fluorophores. We support our findings with histograms of fluorescence bursts and with an analytical derivation of the interphoton delay distribution of (nearly) immobilized emitters from the fluorescence intensity profile.

Gold Nanoparticles as Absolute Nanothermometers.

Nanothermometry is a challenging field that can open the door to intriguing questions ranging from biology and medicine to material sciences. Gold nanorods are excellent candidates to act as nanoprobes because they are reasonably bright emitters upon excitation with a monochromatic source. Gold nanoparticles are commonly used in photothermal therapy as efficient transducers of electromagnetic radiation into heat. In this work we use the spectrum of the anti-Stokes emission from gold nanorods irradiated in resonance to measure the absolute temperature of the nanoparticles and their surrounding medium without the need for a previous calibration. We show a 4 K accuracy in the determination of the temperature of the medium with spectral measurements of 180 s integration time. This procedure can be easily implemented in any microscope capable of acquiring emission spectra, and it is not limited to any specific shape of nanoparticles.

Gold Nanorod Enhanced Fluorescence Enables Single-Molecule Electrochemistry of Methylene Blue.

Redox reactions are central to energy conversion and life metabolism. Herein we present electrochemical measurements with fluorescent readout of the redox-sensitive dye Methylene Blue (MB), at the single-molecule (SM) level. To overcome the low fluorescence quantum yield of MB we enhanced fluorescence by using individual gold nanorods to achieve the required sensitivity. By measuring the same molecule at different electrochemical potentials we determined the mid-point potential of each single molecule through its redox-induced fluorescence blinking dynamics.

Monitoring in real-time focal adhesion protein dynamics in response to a discrete mechanical stimulus.

The adhesion of cells to the extracellular matrix is a hierarchical, force-dependent, multistage process that evolves at several temporal scales. An understanding of this complex process requires a precise measurement of forces and its correlation with protein responses in living cells. We present a method to quantitatively assess live cell responses to a local and specific mechanical stimulus. Our approach combines atomic force microscopy with fluorescence imaging. Using this approach, we evaluated the recruitment of adhesion proteins such as vinculin, focal adhesion kinase, paxillin, and zyxin triggered by applying forces in the nN regime to live cells. We observed in real time the development of nascent adhesion sites, evident from the accumulation of early adhesion proteins at the position where the force was applied. We show that the method can be used to quantify the recruitment characteristic times for adhesion proteins in the formation of focal complexes. We also found a spatial remodeling of the mature focal adhesion protein zyxin as a function of the applied force. Our approach allows the study of a variety of complex biological processes involved in cellular mechanotransduction.

Non-plasmonic nanoantennas for surface enhanced spectroscopies with ultra-low heat conversion.

Nanoplasmonics has recently revolutionized our ability to control light on the nanoscale. Using metallic nanostructures with tailored shapes, it is possible to efficiently focus light into nanoscale field 'hot spots'. High field enhancement factors have been achieved in such optical nanoantennas, enabling transformative science in the areas of single molecule interactions, highly enhanced nonlinearities and nanoscale waveguiding. Unfortunately, these large enhancements come at the price of high optical losses due to absorption in the metal, severely limiting real-world applications. Via the realization of a novel nanophotonic platform based on dielectric nanostructures to form efficient nanoantennas with ultra-low light-into-heat conversion, here we demonstrate an approach that overcomes these limitations. We show that dimer-like silicon-based single nanoantennas produce both high surface enhanced fluorescence and surface enhanced Raman scattering, while at the same time generating a negligible temperature increase in their hot spots and surrounding environments.

High-efficiency second harmonic generation from a single hybrid ZnO nanowire/Au plasmonic nano-oligomer.

We introduce a plasmonic-semiconductor hybrid nanosystem, consisting of a ZnO nanowire coupled to a gold pentamer oligomer by crossing the hot-spot. It is demonstrated that the hybrid system exhibits a second harmonic (SH) conversion efficiency of ∼3 × 10(-5)%, which is among the highest values for a nanoscale object at optical frequencies reported so far. The SH intensity was found to be ∼1700 times larger than that from the same nanowire excited outside the hot-spot. Placing high nonlinear susceptibility materials precisely in plasmonic confined-field regions to enhance SH generation opens new perspectives for highly efficient light frequency up-conversion on the nanoscale.