Excitation Spot Research Articles

Coherent-hybrid STED: high contrast sub-diffraction imaging using a bi-vortex depletion beam.

Stimulated emission depletion (STED) fluorescence microscopy squeezes an excited spot well below the wavelength scale using a doughnut-shaped depletion beam. To generate a doughnut, a scale-free vortex phase modulation (2D-STED) is often used because it provides maximal transverse confinement and radial-aberration immunity (RAI) to the central dip. However, RAI also means blindness to a defocus term, making the axial origin of fluorescence photons uncertain within the wavelength scale provided by the confocal detection pinhole. Here, to reduce the uncertainty, we perturb the 2D-STED phase mask so as to change the sign of the axial concavity near focus, creating a dilated dip. By providing laser depletion power, the dip can be compressed back in three dimensions to retrieve lateral resolution, now at a significantly higher contrast. We test this coherent-hybrid STED (CH-STED) mode in x-y imaging of complex biological structures, such as the dividing cell. The proposed strategy creates an orthogonal direction in the STED parametric space that uniquely allows independent tuning of resolution and contrast using a single depletion beam in a conventional (circular polarization-based) STED setup.

Spatial distribution of the luminescence of an electron-hole system in a double-well Si/SiGe/Si heterostructure

The in-plane spatial distribution of photoluminescence (PL) intensity in two-dimensional layers of a type-II (buffer Si1-yGey)/tSi/sSi1-xGex/tSi/(cap Si1-yGey) heterostructure is investigated at liquid-helium temperatures at high excitation levels. It is found that the PL spectra recorded at the excitation spot and the edge of the sample are different. This work examines the dependences of the PL spectra and intensity both on the distance between the observation point and a fixed excitation spot and on the distance between the excitation spot and a fixed observation point at the edge of the sample. The reasons for the observed behavior of the PL are determined.

Photon-separation to enhance the spatial resolution of pulsed STED microscopy.

Stimulated emission depletion microscopy (STED) is one of the pivotal super-resolution techniques. It overcomes the spatial resolution limit imposed by the diffraction by using an additional laser beam, the STED beam, intensity of which is directly related to the achievable resolution. Despite reaching nanometer resolution, much effort in recent years has been devoted to reducing the STED beam intensity because it may lead to photo-damaging effects. Accessing the spatial information encoded in the temporal dynamics of the detected fluorescent photons has been proved to be a powerful strategy and has contributed to the separation by lifetime tuning (SPLIT) technique. The SPLIT method uses the phasor analysis to efficiently distinguish photons emitted from the center and the periphery of the excitation spot. It thus improves the resolution without increasing the STED beam intensity. This method was proposed for architectures based on the STED beam running in continuous waves (CW-STED microscopy). Here, we extend it to pulsed STED beam implementations (pSTED microscopy). We show, through simulated and experimental data, that the pSTED-SPLIT method reduces the detection volume of the pSTED microscope without significantly decreasing the signal-to-noise ratio of the final image, thus effectively improving the resolution without increasing the STED beam intensity.

Spatiotemporal Evolution of Coherent Polariton Modes in ZnO Microwire Cavities at Room Temperature

Tunable waveguides for propagating coherent quantum states are demanded for future applications in quantum information technology and optical data processing. We present coherent whispering gallery mode polariton states in ZnO-based hexagonal microwires at room temperature. We observed their propagation over the field of view of about 20 μm by picosecond time-resolved real space imaging using a streak camera. Spatial coherence was proven by time integrated Michelson interferometry superimposing the inverted spatial emission pattern with its original one. We furthermore show that the real and momentum space evolution of the coherent states can not only be described by the commonly used model developed for ballistically propagating Bose-Einstein condensates based on the Gross-Pitaevskii equation but equivalently by classical ray optics considering a spatially varying particle density dependent refractive index of the cavity material, not yet considered in literature so far. By changing the excitation spot size, the refractive index gradient and thus the propagation velocity is changed.

Enhancement of fluorescence emission difference microscopy using conjugated vortex phase modulation.

In this paper, we propose and demonstrate an improved fluorescence emission difference microscopy (FED) method, exploiting programmable phase modulation for image enhancement. The main novelty of the proposed approach lies in the matched size and intensity of two excitation spots. The proposed method improves the FED performance on image quality via artefact elimination. We demonstrate the feasibility of this method through theoretical studies and experimental tests. The experimental results of nanobeads and cells validate the practical performance of this method, which can enable reliable observations at superresolution in biomedical studies. LAY DESCRIPTION: In this paper, we propose a method to improve the imaging quality of regular fluorescence emission difference (FED) microscopy. In regular FED imaging, a solid and a doughnut excitation beam are successively used to acquire two images which are then subtracted with each other to improve the resolution of confocal microscopy. The doughnut beam can be generated by modulating the excitation beam with a vortex phase mask. Note that both of the excitation beam and the vortex phase mask must have the same handed direction in regular FED microscopy. However, some negative values may be produced and some information may be lost due to the subtraction process in regular FED imaging, which is mainly caused by the mismatched size and intensity of these two excitation spots. To address this issue, we propose conjugated FED (cFED) microscopy which additionally uses a conjugated vortex phase mask to modulate the solid beam to extend its focal spot size to be matched with the doughnut spot, which means the handed direction of the solid beam and the vortex phase mask is different. Besides, in order not to damage the resolution, the doughnut beam needs to be saturated to some degree. The experiment results show that, at the same resolution level, the negative values and the information loss in cFED image are all less than that of regular FED image.

Shifted excitation resonance Raman difference spectroscopy system suitable for the quantitative in vivo detection of carotenoids in human skin

A system for shifted excitation resonance Raman spectroscopy (SERRDS) suitable for the application in medical practice for the in vivo detection of carotenoids in human skin is presented. This system comprises a miniaturized (150 mm × 27 mm × 12 mm) handheld probe and a wavelength-tunable diode laser-based 488 nm SHG light source. The diode laser provides two closely spaced excitation wavelengths. In parallel with the resonance excitation of carotenoids in the skin, SERRDS separates the fluorescence background from the Raman peaks. Inhomogeneities of human skin are averaged by the applied spot diameter of 3 mm. The implemented optics are designed for a detection of carotenoids over the whole excitation spot area. The system was calibrated using skin phantoms, resulting in a detection limit of 0.03 nmol g−1 (beta-carotene per gram of skin/tissue) which is more than one order of magnitude below the average beta-carotene concentration in human skin.

Coupled exciton-trion spin dynamics in a MoSe2 monolayer

Understanding and controlling the spin degree of freedom in two-dimensional transition metal dichalcogenides offers the potential for designing functional quantum materials. This work investigates the dynamics of photo- and resident carrier spins in an encapsulated MoSe2 monolayer using non-degenerate time-resolved Kerr-rotation microscopy. The lightly doped monolayer exhibits clear exciton and trion resonances with spin-polarizations that are characterized by a fast (~20 ps) decay attributed to photocarrier relaxation and recombination, followed by a slow (~690 ps) decay associated with resident carrier depolarization. Dual-frequency Kerr-rotation spectra directly reveal exciton-trion coupling on ultrashort timescales and within the spin coherence time of the system. Moreover, the distribution of the exciton-trion coupling features exposes inhomogeneous broadening likely arising from different domains within the excitation spot.

Phase-sensitive detection of acoustically stimulated electromagnetic response in steel

The signal amplitude and the phase of acoustically stimulated electromagnetic (ASEM) response have been investigated in steel. In the ASEM method, magnetization is temporally modulated with the radio frequency (rf) of irradiated ultrasonic waves through magnetomechanical coupling. The first-harmonic components of the induced rf dipolar magnetic fields are detected using a resonant loop antenna. The signal amplitude of ASEM waves is determined by the magnitude of local piezomagnetic coefficients on an acoustically excited spot. Here, we divided the ASEM waves into the “in-phase” and “quadrature” components by phase-sensitive detection (PSD). On the basis of the linear response theory, we provided the theoretical formalism of ASEM response by introducing local complex piezomagnetic coefficients, dloc = d′ + id′′. We investigated the magnetic field (H) dependence of the individual components on the different surface conditions of steel plates. The in-phase component [∝ d′(H)] shows a hysteresis loop on the machined surface of a steel plate, in which d′(H) switches sign at two finite field values, ±H0. The inversion of magnetization associated with the applied static fields is thus definitely observed in the PSD measurements. In addition, we measured the hysteresis behaviors on a steel surface with a thin mill scale (iron oxide layers). The hysteresis loop broadens and a significant contribution of the quadrature component [∝ d′′(H)] is found. We discuss the origin of the hysteresis behaviors of d′ and d′′ using the Debye relaxation model.

Localization-Delocalization Transition in Disordered One-Dimensional Exciton-Polariton System

The transition from the delocalized to the localized state has been investigated in a quasi-onedimensional exciton-polariton system excited nonresonantly in GaAs-based microcavity wire with disordered potential. The photoexcited polariton condensate has been found to spread along the wire with а velocity exceeding 1 μm/ps. The propagation along the wire is provided by high energy polaritons. The LP localization length decreases with decreasing blue shift of LPs in the excited spot. The polariton condensate returns to the Bose glass state when the blue shift of the LP resonance at the excitation spot decreases below the critical level that depends on the potential disorder.

Recent research on stimulated emission depletion microscopy for reducing photobleaching.

Stimulated emission depletion (STED) microscopy is a useful tool in investigation for super-resolution realm. By silencing the peripheral fluorophores of the excited spot, leaving only the very centre zone vigorous for fluorescence, the effective point spread function (PSF) could be immensely squeezed and subcellular structures, such as organelles, become discernable. Nevertheless, because of the low cross-section of stimulated emission and the short fluorescence lifetime, the depletion power density has to be extremely higher than the excitation power density and molecules are exposed in high risk of photobleaching. The existence of photobleaching greatly limits the research of STED in achieving higher resolution and more delicate imaging quality, as well as long-term and dynamic observation. Since the first experimental implementation of STED microscopy, researchers have lift out variety of methods and techniques to alleviate the problem. This paper would present some researches via conventional methods which have been explored and utilised relatively thoroughly, such as fast scanning, time-gating, two-photon excitation (TPE), triplet relaxation (T-Rex) and background suppression. Alternatively, several up-to-date techniques, especially adaptive illumination, would also be unveiled for discussion in this paper. The contrast and discussion of these modalities would play an important role in ameliorating the research of STED microscopy.

Active control of radiation beaming from Tamm nanostructures by optical microscopy

Active control of the radiation orientation (beaming) of a metallic antenna has been reported by various methods, where the antenna excitation position was tuned with a typical 50 nm precision by a near-field tip or an electron-beam. Here we use optical microscopy to excite and analyze the fluorescence of a layer of nanocrystals embedded in an optical Tamm state nanostructure (metallic disk on top of a Bragg mirror). We show that the radiation pattern can be controlled by changing the excitation spot on the disk with only micrometer precision, in a manner which can be well described by numerical simulations. A simplified analytical model suggests that the propagation length of the in-plane confined optical modes is a key parameter for beaming control.

Localization-delocalization transition in disordered one-dimensional exciton-polariton system

AbstractThe transition from the delocalized to the localized state has been investigated in a quasi-onedimensional exciton-polariton system excited nonresonantly in GaAs-based microcavity wire with disordered potential. The photoexcited polariton condensate has been found to spread along the wire with а velocity exceeding 1 μm/ps. The propagation along the wire is provided by high energy polaritons. The LP localization length decreases with decreasing blue shift of LPs in the excited spot. The polariton condensate returns to the Bose glass state when the blue shift of the LP resonance at the excitation spot decreases below the critical level that depends on the potential disorder.

Single shot all optical switching of intrinsic micron size magnetic domains of a Pt/Co/Pt ferromagnetic stack

We demonstrate that magnetization reversal in a ferromagnetic Pt/Co/Pt stack can be induced by a single femtosecond laser pulse. We find that the size of the switched spot is comparable to the size of the intrinsic magnetic domains. It requires an absorbed energy density of ∼4 mJ cm−2, beyond which the excited spot fragments into a multidomain structure. The switching process can be toggled with sequential laser pulses and is helicity-independent. Furthermore, the dynamics of the magnetization reversal occurs in a timescale less than one microsecond. These results suggest that all-optical switching in ferromagnetic films requires that the laser spot matches with the specific domain sizes.

Three-dimensional weight-accumulation algorithm for generating multiple excitation spots in fast optical stimulation

We report here an algorithm for calculating a hologram to be employed in a high-access speed microscope for observing sensory-driven synaptic activity across all inputs to single living neurons in an intact cerebral cortex. The system is based on holographic multi-beam generation using a two-dimensional phase-only spatial light modulator to excite multiple locations in three dimensions with a single hologram. The hologram was calculated with a three-dimensional weighted iterative Fourier transform method using the Ewald sphere restriction to increase the calculation speed. Our algorithm achieved good uniformity of three dimensionally generated excitation spots; the standard deviation of the spot intensities was reduced by a factor of two compared with a conventional algorithm.

Optical crosstalk in SPAD arrays for high-throughput single-molecule fluorescence spectroscopy

Single-molecule fluorescence spectroscopy (SMFS), based on the detection of individual molecules freely diffusing through the excitation spot of a confocal microscope, has allowed unprecedented insights into biological processes at the molecular level, but suffers from limited throughput. We have recently introduced a multispot version of SMFS, which allows achieving high-throughput SMFS by virtue of parallelization, and relies on custom silicon single-photon avalanche diode (SPAD) detector arrays. Here, we examine the premise of this parallelization approach, which is that data acquired from different spots is uncorrelated. In particular, we measure the optical crosstalk characteristics of the two 48-pixel SPAD arrays used in our recent SMFS studies, and demonstrate that it is negligible (crosstalk probability ≤1.110−3) and undetectable in cross-correlation analysis of actual single-molecule fluorescence data.

48-spot single-molecule FRET setup with periodic acceptor excitation.

Single-molecule Förster resonance energy transfer (smFRET) allows measuring distances between donor and acceptor fluorophores on the 3-10 nm range. Solution-based smFRET allows measurement of binding-unbinding events or conformational changes of dye-labeled biomolecules without ensemble averaging and free from surface perturbations. When employing dual (or multi) laser excitation, smFRET allows resolving the number of fluorescent labels on each molecule, greatly enhancing the ability to study heterogeneous samples. A major drawback to solution-based smFRET is the low throughput, which renders repetitive measurements expensive and hinders the ability to study kinetic phenomena in real-time. Here we demonstrate a high-throughput smFRET system that multiplexes acquisition by using 48 excitation spots and two 48-pixel single-photon avalanche diode array detectors. The system employs two excitation lasers allowing separation of species with one or two active fluorophores. The performance of the system is demonstrated on a set of doubly labeled double-stranded DNA oligonucleotides with different distances between donor and acceptor dyes along the DNA duplex. We show that the acquisition time for accurate subpopulation identification is reduced from several minutes to seconds, opening the way to high-throughput screening applications and real-time kinetics studies of enzymatic reactions such as DNA transcription by bacterial RNA polymerase.

2D magnetofermionic condensate in GaAs/AlGaAs heterostructures

A principally new collective state—magnetofermionic condensate—was found upon excitation of long-lived triplet cyclotron magnetoexcitons in a Hall insulator with high electron mobility and filling factor ν = 2 at low temperatures T < 1 K. The condensed phase interacts coherently with external electromagnetic field, exhibits superradiant properties and spreads over macroscopic distances from the excitation spot along the surface of the 2D structure due to its low viscosity. The observed effects are explained in terms of a coherent condensate arising in the non-equilibrium system of 2D fermions with a fully quantized energy spectrum, in which a degenerate ensemble of long-lived triplet magnetoexcitons with Bose statistics is formed. The condensation occurs in the space of magnetic translation vectors. Under these conditions, new opportunities arise for future physical studies.

Generation of spin waves by fs-laser pulses in transparent magnetic films: role of the laser beam diameter

Magnetostatic spin waves (MSWs) in magnetic dielectrics are perspective candidates for various applications including data processing, spin wave logic and quantum information. They can propagate hundreds of microns, some have nonreciprocal character and can be localized near the film interfaces. Excitation of MSWs by fs-laser pulses is beneficial since it provides local excitation and tunability. We demonstrate a method for the optical pumping of MSWs due to the inverse Faraday effect in magnetic dielectrics. Its essence is in the excitation with a nonuniform distribution of the optical power through the film depth and in optimization of the pump beam size. Propagation of the spin waves is demonstrated by detecting the magnetization dynamics tens microns away from the excitation spot. Variation of the pump beam diameter allows tuning between different types of the excited spin waves.

Spatially resolved and time-resolved imaging of transport of indirect excitons in high magnetic fields

We present the direct measurements of magnetoexciton transport. Excitons give the opportunity to realize the high magnetic field regime for composite bosons with magnetic fields of a few Tesla. Long lifetimes of indirect excitons allow the study kinetics of magnetoexciton transport with time-resolved optical imaging of exciton photoluminescence. We performed spatially, spectrally, and time-resolved optical imaging of transport of indirect excitons in high magnetic fields. We observed that increasing magnetic field slows down magnetoexciton transport. The time-resolved measurements of the magnetoexciton transport distance allowed for an experimental estimation of the magnetoexciton diffusion coefficient. An enhancement of the exciton photoluminescence energy at the laser excitation spot was found to anti-correlate with the exciton transport distance. A theoretical model of indirect magnetoexciton transport is presented and is in agreement with the experimental data.

Macroscopic Two-Dimensional Polariton Condensates.

We report a record-size, two-dimensional polariton condensate of a fraction of a millimeter radius free from the presence of an exciton reservoir. This macroscopically occupied state is formed by the ballistically expanding polariton flow that relaxes and condenses over a large area outside of the excitation spot. The density of this trap-free condensate is <1 polariton/μm^{2}, reducing the phase noise induced by the interaction energy. Moreover, the backflow effect, recently predicted for the nonparabolic polariton dispersion, is observed here for the first time in the fast-expanding wave packet.