Two-photon excited lasing for detection of amyloids in brain tissue.

Two-photon excitation of emissive markers with near-infrared (NIR) light is of a particular interest for imaging in biology and medicine because NIR light is relatively weakly absorbed and scattered by tissues. At the same time the mechanism of two-photon absorption allows excitation of molecules located deep inside a scattering medium. In this work we demonstrate that the two-photon excitation combined with the effect of light amplification in the stimulated emission process provides a sensitive method for detecting amyloids of different forms. We investigate the two-photon excited amplified spontaneous emission (ASE) of a fluorescent dye, coumarin 307, in the brain tissue infiltrated with various amyloid phantoms i.e. oligomers, protofibrils and mature fibrils. All these forms of amyloids can be detected by observation of ASE and determination of thresholds for light amplification. On this basis we suggest that a relatively simple extension of currently used emission-based optical spectroscopy techniques can provide key information on pathogenic amyloid structures in tissue.

First events in the coil-to-globule transition of PVME in water: An ultrafast temperature jump - time-resolved elastic light scattering study.

HYPOTHESIS: The coil-to-globule transition is an essential phenomenon in protein and polymer solutions. Late stages of such transitions, >1 µs, have been thoroughly studied. Yet, the initial ones are a matter of speculations. Here, we present the first observation of a sub-nanosecond stage of the coil-to-globule transition of poly (vinyl methyl ether), PVME, in water. EXPERIMENTS: The detection of an early stage of the coil-to-globule transition has been possible thanks to a novel experimental approach - time-resolved elastic light scattering study, following an ultrafast temperature jump. We identified a molecular process active in the observed stage of the transition with use of broadband dielectric spectroscopy. FINDINGS: In the experiment's time window, from a few ps to around 600 ps, we observed an increase in the light scattering intensity 300-400 ps after the temperature jump that heated the sample above its lower critical solution temperature (LCST). The observed time coincides with the time of segmental relaxation of PVME, determined by broadband dielectric spectroscopy in the temperature range of the LCST of the PVME/water mixture. This coincidence strongly suggests that the observed herein stage of coil-to-globule transition is the rapid formation of local nuclei along the polymer chain. Those nuclei may grow and aggregate in later stages of the process, which are out of our experimental time window.

Spontaneous versus Stimulated Surface-Enhanced Raman Scattering of Liquid Water.

We have observed for the first time the surface-enhanced (SE) signal of water in an aqueous dispersion of silver nanoparticles in spontaneous (SERS) and femtosecond stimulated Raman (SE-FSRS) processes with different wavelengths of the Raman pump (515, 715, and 755 nm). By estimating the fraction of water molecules that interact with the metal surface, we have calculated enhancement factors (EF): 4.8 × 106 for SERS and (3.6-3.7) × 106 for SE-FSRS. Furthermore, we have tested the role of simultaneous plasmon resonance and Raman resonance conditions for the aν 1 + bν3 overtone mode of water (755 nm) in SE-FSRS signal amplification. When the wavelength of the Raman pump is within the plasmon resonance of the metal nanoparticles, the Raman resonance has a negligible effect on the EF. However, the Raman resonance with the aν 1 + bν3 mode strongly enhances the signal of the fundamental OH stretching mode of water.

Low noise, self-referenced all polarization maintaining Ytterbium fiber laser frequency comb: erratum.

We provide a corrected figure of our previous publication [Opt. Express25, 18017 (2017)10.1364/OE.25.018017].

Influence of local microenvironment on the double hydrogen transfer in porphycene.

We performed time-resolved transient absorption and fluorescence anisotropy measurements in order to study tautomerization of porphycene in rigid polymer matrices at cryogenic temperatures. Studies were carried out in poly(methyl methacrylate) (PMMA), poly(vinyl butyral) (PVB), and poly(vinyl alcohol) (PVA). The results prove that in all studied media hydrogen tunnelling plays a significant role in the double hydrogen transfer which becomes very sensitive to properties of the environment below approx. 150 K. We also demonstrate that there exist two populations of porphycene molecules in rigid media: "hydrogen-transferring" molecules, in which tautomerization occurs on time scales below 1 ns and "frozen" molecules in which double hydrogen transfer is too slow to be monitored with nanosecond techniques. The number of "frozen" molecules increases when the sample is cooled. We explain this effect by interactions of guest molecules with a rigid host matrix which disturbs symmetry of porphycene and hinders tunnelling. Temperature dependence of the number of hydrogen-transferring molecules suggests that the factor which restores the symmetry of the double-minimum potential well in porphycene are intermolecular vibrations localized in separated regions of the amorphous polymer.

Raman-induced pulse destabilization and bistability in an all-normal dispersion oscillator.

We report the experimental study of spectral modulations induced by a stimulated Raman scattering process in an all-fiber all-normal dispersion oscillator. With the use of dispersive Fourier transform, we recorded a series of single-shot spectra of consecutive laser pulses. The data indicate that the Raman process destabilizes the long-wavelength part of the laser pulse spectrum without disrupting the single-pulse generation regime. Our experiments revealed also that the oscillator displayed bistable operation for high pump powers. Two stable dissipative soliton mode-locked states were observed, together with output power hysteresis.

Femtosecond infrared pump-stimulated Raman probe spectroscopy: the first application of the method to studies of vibrational relaxation pathways in the liquid HDO/D2O system.

We have proposed and constructed a setup for a novel method of ultrafast vibrational spectroscopy: femtosecond infrared pump-stimulated Raman probe spectroscopy. This is the first time-resolved spectroscopy providing simultaneously a sub-100 fs time resolution, a spectral resolution better than 10 cm-1 and a spectral window covering an extremely broad range of molecular vibrations (at least: 200-4000 cm-1) with a "single laser shot". The new method was applied to study vibrational relaxation pathways in the liquid HDO/D2O system. We determined the lifetimes of OH stretching vibrations to be in the range 310-500 fs depending on the isotopic dilution, which is in good agreement with the results from pump-probe femtosecond infrared spectroscopy. Moreover, we observed a strong coupling of OH stretch to OD stretch vibrations and possibly also to the librational modes of water.

Nonlinear refractive index measurement by SPM-induced phase regression.

Herewith, we describe how intensity and phase of the ultrashort pulse retrieved with second-harmonic frequency-resolved optical gating (SHG FROG) can be utilized for measurement of the nonlinear refractive index (n 2). Through comparison with available literature, we show that our method surpasses Z-scan in terms of precision by a factor of two, and thus, constitutes an interesting alternative. We present results for various materials: fused silica, calcite, YVO 4, BiBO, CaF 2, and YAG at 1030 nm. Unlike the Z-scan, the use of this method is not restricted to free-space geometry, but due to its characteristics, it can be used in integrated waveguides or photonic crystal fibers as well.

Two-photon excited lasing of Coumarin 307 for lysozyme amyloid fibrils detection.

Amyloid fibrils are a well-recognized hallmark of neurodegeneration. A common approach to detect amyloid fibrils is staining with organic molecules and monitoring optical properties using fluorescence spectroscopy. However, the structural diversity of amyloids necessitates new sensitive methods and probes that can be reliably used to characterize them. Here, Coumarin 307 is applied for lysozyme fibrils detection by observation of laser action in the process of two-photon excited stimulated emission. It is shown that the lasing threshold and spectrum significantly depend on the adopted structure (α-helix or ß-sheet) of the lysozyme protein, whereas fluorescence spectrum is insensitive to the protein structure. The applications of coherent stimulated emission light that can be emitted deep inside a scattering medium can be particularly promising for imaging and therapeutic purposes in the neurodegeneration field. Two-photon excitation with the near-infrared light, which allows the deepest penetration of tissues, is an important advantage of the method.

Noncollinear and nonlinear pulse propagation.

A novel method for numerical modelling of noncollinear and nonlinear interaction of femtosecond laser pulses is presented. The method relies on a separate treatment of each of the interacting pulses by it's own rotated unidirectional pulse propagation equation (UPPE). We show that our method enables accurate simulations of the interaction of pulses travelling at a mutual angle of up to 140°. The limit is imposed by the unidirectionality principal. Additionally, a novel tool facilitating the preparation of noncollinear propagation initial conditions - a 3D Fourier transform based rotation technique - is presented. The method is tested with several linear and nonlinear cases and, finally, four original results are presented: (i) interference of highly chirped pulses colliding at mutual angle of 120°, (ii) optical switching through cross-focusing of perpendicular beams (iii) a comparison between two fluorescence up-conversion processes in BBO with large angles between the input beams and (iv) a degenerate four-wave mixing experiment in a boxcar configuration.

Nonlinear polarization evolution of ultrashort pulses in polarization maintaining fibers.

We examine properties of an ultrashort laser pulse propagating through an artificial Saturable Absorber based on Nonlinear Polarization Evolution device which has been realized with Polarization Maintaining fibers only (PM NPE). We study and compare in-line and Faraday Mirror geometries showing that the latter is immune to errors in the PM NPE construction. Experimental results for the transmission measurements of the PM NPE setup for different input linear polarization angles and various input pulse powers are presented. We show that PM NPE topology is of crucial importance for controlling the properties of the output pulse as it rules the contribution of cross-phase modulation to an overall nonlinear phase change. We also demonstrate an excellent agreement between the numerical model and experimental results.

Photobleaching in STED nanoscopy and its dependence on the photon flux applied for reversible silencing of the fluorophore.

In STED (stimulated emission depletion) nanoscopy, the resolution and signal are limited by the fluorophore de-excitation efficiency and photobleaching. Here, we investigated their dependence on the pulse duration and power of the applied STED light for the popular 750 nm wavelength. In experiments with red- and orange-emitting dyes, the pulse duration was varied from the sub-picosecond range up to continuous-wave conditions, with average powers up to 200 mW at 80 MHz repetition rate, i.e. peak powers up to 1 kW and pulse energies up to 2.5 nJ. We demonstrate the dependence of bleaching on pulse duration, which dictates the optimal parameters of how to deliver the photons required for transient fluorophore silencing. Measurements with the dye ATTO647N reveal that the bleaching of excited molecules scales with peak power with a single effective order ~1.4. This motivates peak power reduction while maintaining the number of STED-light photons, in line with the superior resolution commonly achieved for nanosecond STED pulses. Other dyes (ATTO590, STAR580, STAR635P) exhibit two distinctive bleaching regimes for constant pulse energy, one with strong dependence on peak power, one nearly independent. We interpret the results within a photobleaching model that guides quantitative predictions of resolution and bleaching.

Low noise, self-referenced all polarization maintaining Ytterbium fiber laser frequency comb.

We report the implementation of a self-referenced optical frequency comb generated by a passively mode-locked all polarization maintaining (PM) Yb fiber laser based on a nonlinear amplifying loop mirror (NALM). After spectral broadening the optical spectrum spans from 650 nm to 1400 nm, allowing for the generation of an optical octave and carrier envelope offset frequency (fceo) stabilization through a conventional f-2f interferometer. We demonstrate for the first time the stabilization of the fceo of such a PM Yb system with an in-loop fractional frequency stability scaled to an optical frequency of low 10-19 at 1 second averaging time, offering a great potential for applications in optical atomic clock metrology.

How good is the generalized Langevin equation to describe the dynamics of photo-induced electron transfer in fluid solution?

The dynamics of unimolecular photo-triggered reactions can be strongly affected by the surrounding medium for which a large number of theoretical descriptions have been used in the past. An accurate description of these reactions requires knowing the potential energy surface and the friction felt by the reactants. Most of these theories start from the Langevin equation to derive the dynamics, but there are few examples comparing it with experiments. Here we explore the applicability of a Generalized Langevin Equation (GLE) with an arbitrary potential and a non-Markovian friction. To this end, we have performed broadband fluorescence measurements with sub-picosecond time resolution of a covalently linked organic electron donor-acceptor system in solvents of changing viscosity and dielectric permittivity. In order to establish the free energy surface (FES) of the reaction, we resort to stationary electronic spectroscopy. On the other hand, the dynamics of a non-reacting substance, Coumarin 153, provide the calibrating tool for the non-Markovian friction over the FES, which is assumed to be solute independent. A simpler and computationally faster approach uses the Generalized Smoluchowski Equation (GSE), which can be derived from the GLE for pure harmonic potentials. Both approaches reproduce the measurements in most of the solvents reasonably well. At long times, some differences arise from the errors inherited from the analysis of the stationary solvatochromism and at short times from the excess excitation energy. However, whenever the dynamics become slow, the GSE shows larger deviations than the GLE, the results of which always agree qualitatively with the measured dynamics, regardless of the solvent viscosity or dielectric properties. The method applied here can be used to predict the dynamics of any other reacting system, given the FES parameters and solvent dynamics are provided. Thus no fitting parameters enter the GLE simulations, within the applicability limits found for the model in this work.

Ground State Depletion Nanoscopy Resolves Semiconductor Nanowire Barcode Segments at Room Temperature.

Nanowires hold great promise as tools for probing and interacting with various molecular and biological systems. Their unique geometrical properties (typically <100 nm in diameter and a few micrometers in length) enable minimally invasive interactions with living cells, so that electrical signals or forces can be monitored. All such experiments require in situ high-resolution imaging to provide context. While there is a clear need to extend visualization capabilities to the nanoscale, no suitable super-resolution far-field photoluminescence microscopy of extended semiconductor emitters has been described. Here, we report that ground state depletion (GSD) nanoscopy resolves heterostructured semiconductor nanowires formed by alternating GaP/GaInP segments ("barcodes") at a 5-fold resolution enhancement over confocal imaging. We quantify the resolution and contrast dependence on the dimensions of GaInP photoluminescence segments and illustrate the effects by imaging different nanowire barcode geometries. The far-red excitation wavelength (∼700 nm) and low excitation power (∼3 mW) make GSD nanoscopy attractive for imaging semiconductor structures in biological applications.

Full 3D modelling of pulse propagation enables efficient nonlinear frequency conversion with low energy laser pulses in a single-element tripler.

Although new optical materials continue to open up access to more and more wavelength bands where femtosecond laser pulses can be generated, light frequency conversion techniques are still indispensable in filling the gaps on the ultrafast spectral scale. With high repetition rate, low pulse energy laser sources (oscillators) tight focusing is necessary for a robust wave mixing and the efficiency of broadband nonlinear conversion is limited by diffraction as well as spatial and temporal walk-off. Here we demonstrate a miniature third harmonic generator (tripler) with conversion efficiency exceeding 30%, producing 246 fs UV pulses via cascaded second order processes within a single laser beam focus. Designing this highly efficient and ultra compact frequency converter was made possible by full 3-dimentional modelling of propagation of tightly focused, broadband light fields in nonlinear and birefringent media.

Influence of the excitation light intensity on the rate of fluorescence quenching reactions: pulsed experiments.

The effect of multiple light excitation events on bimolecular photo-induced electron transfer reactions in liquid solution is studied experimentally. It is found that the decay of fluorescence can be up to 25% faster if a second photon is absorbed after a first cycle of quenching and recombination. A theoretical model is presented which ascribes this effect to the enrichment of the concentration of quenchers in the immediate vicinity of fluorophores that have been previously excited. Despite its simplicity, the model delivers a qualitative agreement with the observed experimental trends. The original theory by Burshtein and Igoshin (J. Chem. Phys., 2000, 112, 10930-10940) was created for continuous light excitation though. A qualitative extrapolation from the here presented pulse experiments to the continuous excitation conditions lead us to conclude that in the latter the order of magnitude of the increase of the quenching efficiency upon increasing the light intensity of excitation, must also be on the order of tens of percent. These results mean that the rate constant for photo-induced bimolecular reactions depends not only on the usual known factors, such as temperature, viscosity and other properties of the medium, but also on the intensity of the excitation light.

Ultrafast laser mode-locked using nonlinear polarization evolution in polarization maintaining fibers.

We demonstrate an all-fiber ultrafast ytterbium laser oscillator mode-locked by means of a nonlinear polarization evolution (NPE) method realized in polarization-maintaining (PM) fibers. A sequence of the PM fiber pieces is shown to perform NPE action while maintaining a required temporal overlap of the ordinary and extraordinary pulses propagating through it. We present details of simple numerical simulations showing the advantage of the proposed scheme of segmented PM fibers. The laser utilizing the above mentioned design which generates ultrashort pulses at a 20.54 MHz repetition rate with the dechirped pulse duration around 150 fs and a pulse energy of 0.85 nJ is also presented.

Characterization of dimethylsulfoxide/glycerol mixtures: a binary solvent system for the study of "friction-dependent" chemical reactivity.

The properties of binary mixtures of dimethylsulfoxide and glycerol, measured using several techniques, are reported. Special attention is given to those properties contributing or affecting chemical reactions. In this respect the investigated mixture behaves as a relatively simple solvent and it is especially well suited for studies on the influence of viscosity on chemical reactivity. This is due to the relative invariance of the dielectric properties of the mixture. However, special caution must be taken with specific solvation, as the hydrogen-bonding properties of the solvent change with the molar fraction of glycerol.

Spectral compression of femtosecond pulses using chirped volume Bragg gratings.

In this Letter, we demonstrate a 360 fold spectral bandwidth reduction of femtosecond laser pulses using the method of sum frequency generation of pulses with opposite chirps. The reduction has been achieved in a compact setup in which a single chirped volume Bragg grating replaces conventional stretcher and compressor units. Starting with 180 fs pulses, we have obtained, with a 30% overall efficiency, pulses longer than 100 ps with the spectral bandwidth of 0.23 cm-1 (7 GHz). We also discuss our method on theoretical grounds.