Quantitative Determination of Dark Chromophore Population Explains the Apparent Low Quantum Yield of Red Fluorescent Proteins.

The fluorescence quantum yield of four representative red fluorescent proteins mCherry, mKate2, mRuby2, and the recently introduced mScarlet was investigated. The excited state lifetimes were measured as a function of the distance to a gold mirror in order to control the local density of optical states (LDOS). By analyzing the total emission rates as a function of the LDOS, we obtain separately the emission rate and the nonradiative rate of the bright states. We thus obtain for the first time the bright state quantum yield of the proteins without interference from dark, nonemitting states. The bright state quantum yields are considerably higher than previously reported quantum yields that average over both bright and dark states. We determine that mCherry, mKate2, and mRuby2 have a considerable fraction of dark chromophores up to 45%, which explains both the low measured quantum yields of red emitting proteins reported in the literature and the difficulties in developing high quantum yield variants of such proteins. For the recently developed bright mScarlet, we find a much smaller dark fraction of 14%, accompanied by a very high quantum yield of the bright state of 81%. The presence of a considerable fraction of dark chromophores has implications for numerous applications of fluorescent proteins, ranging from quantitative fluorescence microscopy to FRET studies to monitoring protein expression levels. We recommend that future optimization of red fluorescent proteins should pay more attention to minimizing the fraction of dark proteins.

Multiband modulation spectroscopy for the determination of sex and species of mosquitoes in flight.

We present a dual-wavelength polarimetric measurement method to distinguish species and sexes of disease transmitting mosquitoes in flight. By measuring co- and de-polarized backscattered light at 808 and 1550 nm, the degree of linear polarization, wingbeat frequency, reflectance, spectral ratio and glossiness of mosquitoes can be retrieved. Body and wing contributions to these signals can be separated. Whereas the optical cross section is sensitive to the aspect of observation, thus the heading direction of the insect in flight, we demonstrate that polarimetric- and spectral-band ratios are largely invariant to the aspect of observation. We show that wing glossiness, as well as wing- and body-spectral ratios are particularly efficient in distinguishing Anopheles coluzzii and Anopheles arabiensis, 2 closely related species of malaria vectors. Spectral and polarimetric ratios relate to microstructural and melanization features of the wing and body of these species. We conclude that multiband modulation spectroscopy is a useful expansion of the parameter space that can be used to improve the specificity of entomological lidars.

Optical antennas with sinusoidal modulation in width.

Small metal structures sustaining plasmon resonances in the optical regime are of great interest due to their large scattering cross sections and ability to concentrate light to subwavelength volumes. In this paper, we study the dipolar plasmon resonances of optical antennas with a constant volume and a sinusoidal modulation in width. We experimentally show that by changing the phase of the width-modulation, with a small 10 nm modulation amplitude, the resonance shifts over 160 nm. Using simulations we show how this simple design can create resonance shifts greater than 600 nm. The versatility of this design is further shown by creating asymmetric structures with two different modulation amplitudes, which we experimentally and numerically show to give rise to two resonances. Our results on both the symmetric and asymmetric antennas show the capability to control the localization of the fields outside the antenna, while still maintaining the freedom to change the antenna resonance wavelength. The antenna design we tested combines a large spectral tunability with a small footprint: all the antenna dimensions are factor 7 to 13 smaller than the wavelength, and hold potential as a design element in meta-surfaces for beam shaping.

A phased antenna array for surface plasmons.

Surface plasmon polaritons are electromagnetic waves that propagate tightly bound to metal surfaces. The concentration of the electromagnetic field at the surface as well as the short wavelength of surface plasmons enable sensitive detection methods and miniaturization of optics. We present an optical frequency plasmonic analog to the phased antenna array as it is well known in radar technology and radio astronomy. Individual holes in a thick gold film act as dipolar emitters of surface plasmon polaritons whose phase is controlled individually using a digital spatial light modulator. We show experimentally, using a phase sensitive near-field microscope, that this optical system allows accurate directional emission of surface waves. This compact and flexible method allows for dynamically shaping the propagation of plasmons and holds promise for nanophotonic applications employing propagating surface plasmons.

Atomic-scale confinement of resonant optical fields.

In the presence of matter, there is no fundamental limit preventing confinement of visible light even down to atomic scales. Achieving such confinement and the corresponding resonant intensity enhancement inevitably requires simultaneous control over atomic-scale details of material structures and over the optical modes that such structures support. By means of self-assembly we have obtained side-by-side aligned gold nanorod dimers with robust atomically defined gaps reaching below 0.5 nm. The existence of atomically confined light fields in these gaps is demonstrated by observing extreme Coulomb splitting of corresponding symmetric and antisymmetric dimer eigenmodes of more than 800 meV in white-light scattering experiments. Our results open new perspectives for atomically resolved spectroscopic imaging, deeply nonlinear optics, ultrasensing, cavity optomechanics, as well as for the realization of novel quantum-optical devices.

Electrically connected resonant optical antennas.

Electrically connected resonant optical antennas hold promise for the realization of highly efficient nanoscale electro-plasmonic devices that rely on a combination of electric fields and local near-field intensity enhancement. Here we demonstrate the feasibility of such a concept by attaching leads to the arms of a two-wire antenna at positions of minimal near-field intensity with negligible influence on the antenna resonance. White-light scattering experiments in accordance with simulations show that the optical tunability of connected antennas is fully retained. Analysis of the electric properties demonstrates that in the antenna gaps direct current (DC) electric fields of 10(8) V/m can consistently be achieved and maintained over extended periods of time without noticeable damage.

Atomically flat single-crystalline gold nanostructures for plasmonic nanocircuitry.

Deep subwavelength integration of high-definition plasmonic nanostructures is of key importance in the development of future optical nanocircuitry for high-speed communication, quantum computation and lab-on-a-chip applications. To date, the experimental realization of proposed extended plasmonic networks consisting of multiple functional elements remains challenging, mainly because of the multi-crystallinity of commonly used thermally evaporated gold layers. This can produce structural imperfections in individual circuit elements that drastically reduce the yield of functional integrated nanocircuits. In this paper we demonstrate the use of large (>100 µm(2)) but thin (<80 nm) chemically grown single-crystalline gold flakes that, after immobilization, serve as an ideal basis for focused ion beam milling and other top-down nanofabrication techniques on any desired substrate. Using this methodology we obtain high-definition ultrasmooth gold nanostructures with superior optical properties and reproducible nano-sized features over micrometre-length scales. Our approach provides a possible solution to overcome the current fabrication bottleneck and realize high-definition plasmonic nanocircuitry.