Ultrasensitive Label-Free Nanosensing and High-Speed Tracking of Single Proteins

: Label-free detection, analysis, and rapid tracking of nanoparticles is crucial for future ultrasensitive sensing applications, ranging from understanding of biological interactions to the study of size-dependent classical-quantum transitions. Yet optical techniques to distinguish nanoparticles directly among their background remain challenging. Here we present amplified interferometric scattering microscopy (aiSCAT) as a new all-optical method capable of detecting individual nanoparticles as small as 15 kDa proteins that is equivalent to half a GFP. By balancing scattering and reflection amplitudes the interference contrast of the nanoparticle signal is amplified 1 to 2 orders of magnitude. Beyond high sensitivity, a-iSCAT allows high-speed image acquisition exceeding several hundreds of frames-per-second. We showcase the performance of our approach by detecting single Streptavidin binding events and by tracking single Ferritin proteins at 400 frames-per-second with 12 nm localization precision over seconds. Moreover, due to its extremely simple experimental realization, this advancement finally enables a cheap and routine implementation of label-free all-optical single nanoparticle detection platforms with sensitivity operating at the single protein level.Peer ReviewedPostprint (author's final draft

Advances in nanophotonics: ultrafast & ultrasensitive

In this tutorial on NanoPhotonics recent advances are highlighted with focus on near field optical methods, ultra-fast probing of single molecules and ultra-sensitive detection of individual non-fluorescent nanoparticles

A monopole antenna at optical frequencies: single-molecule near-field measurements

We present a monopole antenna for optical frequencies (~600 THz) and discuss near-field measurements with single fluorescent molecules as a technique to characterize such antennas. The similarities and differences between near-field antenna measurements at optical and radio frequencies are discussed in detail. Two typical antenna properties, polarization selectivity and resonances, are studied for the optical monopole by direct near-field measurements and finite integration technique calculations. The antenna is driven by the local field of a sub-wavelength aperture. This gives rise to a dependence of the antenna response on the orientation of the local field vector, in an analogous way to the polarization selectivity of linear wire antennas. The antenna resonances are studied by varying the antenna length. Typical monopole resonances are demonstrated. The finite conductivity of metals at optical frequencies (in combination with the antenna radius) causes the wavelength of the surface charge density oscillation (surface plasmon polariton) along the antenna to be shortened in comparison to the free space wavelength. As a result, resonances for the optical monopole antenna occur at much shorter relative lengths than for conventional radio monopole antennas\ud \u

Nanorod optical antennas for dipolar transitions

Optical antennas link objects to light. Here, we analyze metal nanorod antennas as cavities with variable reflection coefficients to derive the interaction of dipolar transitions with radiation through the antenna modes. The presented analytical model accurately describes the complete emission process, and is summarized in a phase-matching equation. We show how antenna modes evolve as they become increasingly more bound, i.e. plasmonic. The results illustrate why efficient antennas should not be too plasmonic, and how subradiant even modes can evolve into weakly-interacting dark modes. Our description is valid for the interaction of nanorods with light in general, and is thus widely applicable.Comment: 10 pages, 4 figures, submitte

Resonant plasmonic nanoparticles for multicolor second harmonic imaging

Nanoparticles capable of efficiently generating nonlinear optical signals, like second harmonic generation, are attracting a lot of attention as potential background-free and stable nano-probes for biological imaging. However, second harmonic nanoparticles of different species do not produce readily distinguishable optical signals, as the excitation laser mainly defines their second harmonic spectrum. This is in marked contrast to other fluorescent nano-probes like quantum dots that emit light at different colors depending on their sizes and materials. Here, we present the use of resonant plasmonic nanoparticles, combined with broadband phase-controlled laser pulses, as tunable sources of multicolor second harmonic generation. The resonant plasmonic nanoparticles strongly interact with the electromagnetic field of the incident light, enhancing the efficiency of nonlinear optical processes. Because the plasmon resonance in these structures is spectrally narrower than the laser bandwidth, the plasmonic nanoparticles imprint their fingerprints on the second harmonic spectrum. We show how nanoparticles of different sizes produce different colors in the second harmonic spectra even when excited with the same laser pulse. Using these resonant plasmonic nanoparticles as nano-probes is promising for multicolor second harmonic imaging while keeping all the advantages of nonlinear optical microscopy.Peer ReviewedPostprint (published version

Strong antenna-enhanced fluorescence of a single light-harvesting complex shows photon antibunching

The nature of the highly efficient energy transfer in photosynthetic light-harvesting complexes is a subject of intense research. Unfortunately, the low fluorescence efficiency and limited photostability hampers the study of individual light-harvesting complexes at ambient conditions. Here we demonstrate an over 500-fold fluorescence enhancement of light-harvesting complex 2 (LH2) at the single-molecule level by coupling to a gold nanoantenna. The resonant antenna produces an excitation enhancement of circa 100 times and a fluorescence lifetime shortening to ~\n20 ps. The radiative rate enhancement results in a 5.5-fold-improved fluorescence quantum efficiency. Exploiting the unique brightness, we have recorded the first photon antibunching of a single light-harvesting complex under ambient conditions, showing that the 27 bacteriochlorophylls coordinated by LH2 act as a non-classical single-photon emitter. The presented bright antenna-enhanced LH2 emission is a highly promising system to study energy transfer and the role of quantum coherence at the level of single complexes

Ultrafast meets ultrasmall: controlling nanoantennas and molecules

We present a review on the advances of pulse control and ultrafast coherent excitation of both plasmonic nanoantennas and individual molecular systems, primarily based on the achievements in our group. Essential concepts from coherent control of ultrashort broadband laser pulses are combined with nanoscale diffraction limited detection and imaging of single photon emitters; that is, the central area of this work is where ultrafast meets ultrasmall. First, the critical role of dedicated pulse shaping and phase control is discussed, which is crucial to realize free of spatiotemporal coupling Fourier limited pulses inside a high numerical aperture microscope at the diffraction limited spot. Next we apply this scheme to plasmonic antennas, exploiting broadband two-photon excitation, to determine amplitude and phase of plasmonic resonances, to achieve ultrafast switching of nanoscale hotspots, and multicolor second harmonic detection for imaging applications. Subsequently, we address single molecules with phase-shaped pulses to control the electronic state population and retrieve single molecule vibrational dynamics response. We compare the response of a molecule to phase-locked with free phase multipulse excitation. Furthermore, we discuss phase control of excited state energy transfer in photosynthetic molecular complexes. Finally, we combine nanoscale plasmonics with single molecule detection to attain strong enhancement of both excitation and emission, with fluorescence lifetime shortening to the ps range. In conclusion, we anticipate that this review on ultrafast plasmonics and single emitter control will provide a useful view of the status of ultrafast nanophotonics and its application potential.Peer ReviewedPostprint (author's final draft

Control of Vibronic Transition Rates by Resonant Single-Molecule-Nanoantenna Coupling

Plasmonic nanostructures dramatically alter the radiative and nonradiative properties of single molecules in their vicinity. This coupling-induced change in decay channels selectively enhances specific vibronic transitions, which can enable plasmonic control of molecular reactivity. Here, we report coupling-dependent spectral emission shaping of single Rhodamine 800 molecules in the vicinity of plasmonic gold nanorods. We show that the relative vibronic transition rates of the first two vibronic transitions of the spontaneous emission spectrum can be tuned in the weak coupling regime, by approximately 25-fold, on the single molecule level.Peer ReviewedPostprint (author's final draft