A comparative study of CARE 2D and N2V 2D for tissue-specific denoising in second harmonic generation imaging.

This study explored the application of deep learning in second harmonic generation (SHG) microscopy, a rapidly growing area. This study focuses on the impact of glycerol concentration on image noise in SHG microscopy and compares two image restoration techniques: Noise-to-Void 2D (N2V 2D, no reference image restoration) and content-aware image restoration (CARE 2D, full reference image restoration). We demonstrated that N2V 2D effectively restored the images affected by high glycerol concentrations. To reduce sample exposure and damage, this study further addresses low-power SHG imaging by reducing the laser power by 70% using deep learning techniques. CARE 2D excels in preserving detailed structures, whereas N2V 2D maintains natural muscle structure. This study highlights the strengths and limitations of these models in specific SHG microscopy applications, offering valuable insights and potential advancements in the field .

Wave packet dynamics and control in excited states of molecular nitrogen.

Wave packet interferometry with vacuum ultraviolet light has been used to probe a complex region of the electronic spectrum of molecular nitrogen, N2. Wave packets of Rydberg and valence states were excited by using double pulses of vacuum ultraviolet (VUV), free-electron-laser (FEL) light. These wave packets were composed of contributions from multiple electronic states with a moderate principal quantum number (n ∼ 4-9) and a range of vibrational and rotational quantum numbers. The phase relationship of the two FEL pulses varied in time, but as demonstrated previously, a shot-by-shot analysis allows the spectra to be sorted according to the phase between the two pulses. The wave packets were probed by angle-resolved photoionization using an infrared pulse with a variable delay after the pair of excitation pulses. The photoelectron branching fractions and angular distributions display oscillations that depend on both the time delays and the relative phases of the VUV pulses. The combination of frequency, time delay, and phase selection provides significant control over the ionization process and ultimately improves the ability to analyze and assign complex molecular spectra.

Swept coded aperture real-time femtophotography.

Single-shot real-time femtophotography is indispensable for imaging ultrafast dynamics during their times of occurrence. Despite their advantages over conventional multi-shot approaches, existing techniques confront restricted imaging speed or degraded data quality by the deployed optoelectronic devices and face challenges in the application scope and acquisition accuracy. They are also hindered by the limitations in the acquirable information imposed by the sensing models. Here, we overcome these challenges by developing swept coded aperture real-time femtophotography (SCARF). This computational imaging modality enables all-optical ultrafast sweeping of a static coded aperture during the recording of an ultrafast event, bringing full-sequence encoding of up to 156.3 THz to every pixel on a CCD camera. We demonstrate SCARF's single-shot ultrafast imaging ability at tunable frame rates and spatial scales in both reflection and transmission modes. Using SCARF, we image ultrafast absorption in a semiconductor and ultrafast demagnetization of a metal alloy.

Nanoscale reshaping of resonant dielectric microstructures by light-driven explosions.

Femtosecond-laser-assisted material restructuring employs extreme optical intensities to localize the ablation regions. To overcome the minimum feature size limit set by the wave nature of photons, there is a need for new approaches to tailored material processing at the nanoscale. Here, we report the formation of deeply-subwavelength features in silicon, enabled by localized laser-induced phase explosions in prefabricated silicon resonators. Using short trains of mid-infrared laser pulses, we demonstrate the controllable formation of high aspect ratio (>10:1) nanotrenches as narrow as [Formula: see text]. The trench geometry is shown to be scalable with wavelength, and controlled by multiple parameters of the laser pulse train, such as the intensity and polarization of each laser pulse and their total number. Particle-in-cell simulations reveal localized heating of silicon beyond its boiling point and suggest its subsequent phase explosion on the nanoscale commensurate with the experimental data. The observed femtosecond-laser assisted nanostructuring of engineered microstructures (FLANEM) expands the nanofabrication toolbox and opens exciting opportunities for high-throughput optical methods of nanoscale structuring of solid materials.

Complete characterization of a Yb-based OPA at a high repetition rate using frequency resolved optical switching.

We demonstrate experimentally that frequency resolved optical switching (FROSt) can be used to characterize ultra-broadband pulses at high repetition rates up to 500 kHz. Specifically, we present the complete temporal characterization of an optical parametric amplifier (OPA), from the supercontinuum (SC) to the second stage of amplification. Simultaneous characterization of co-propagating signal and idler pulses enables retrieval of their group delay, as well as their temporal phase and intensity. Our study focuses on an extensive frequency range spanning the infrared region (1.2 to 2.4 µm) and confirms the strength and convenience of FROSt as a single tool for characterizing a wide range of pulses at high repetition rates.

Low-energy quasi-circular electron correlations with charge order wavelength in Bi2Sr2CaCu2O8+δ.

Most resonant inelastic x-ray scattering (RIXS) studies of dynamic charge order correlations in the cuprates have focused on the high-symmetry directions of the copper oxide plane. However, scattering along other in-plane directions should not be ignored as it may help understand, for example, the origin of charge order correlations or the isotropic scattering resulting in strange metal behavior. Our RIXS experiments reveal dynamic charge correlations over the qx-qy scattering plane in underdoped Bi2Sr2CaCu2O8+δ. Tracking the softening of the RIXS-measured bond-stretching phonon, we show that these dynamic correlations exist at energies below approximately 70 meV and are centered around a quasi-circular manifold in the qx-qy scattering plane with radius equal to the magnitude of the charge order wave vector, qCO. This phonon-tracking procedure also allows us to rule out fluctuations of short-range directional charge order (i.e., centered around [qx = ±qCO, qy = 0] and [qx = 0, qy = ±qCO]) as the origin of the observed correlations.

Tight-focusing parabolic reflector schemes for petawatt lasers.

A comparative study of three different tight-focusing schemes for high-power lasers is performed numerically. Using the Stratton-Chu formulation, the electromagnetic field in the vicinity of the focus is evaluated for a short-pulse laser beam incident upon an on-axis high numerical aperture parabola (HNAP), an off-axis parabola (OAP), and a transmission parabola (TP). Linearly- and radially-polarized incident beams are considered. It is demonstrated that while all the focusing configurations yield intensities above 1023 W/cm2 for a 1 PW incident beam, the nature of the focused field can be drastically modified. In particular, it is shown that the TP, with its focal point behind the parabola, actually converts an incoming linearly-polarized beam into an m = 2 vector beam. The strengths and weaknesses of each configuration are discussed in the context of future laser-matter interaction experiments. Finally, a generalization of NA calculations up to 4π-illumination is proposed through the solid angle formulation, providing a universal way to compare light cones from any kind of optics.

Mid-infrared frequency domain optical parametric amplifier.

We report on an optical architecture delivering sub-120 femtosecond laser pulses of 20 µJ tunable from 5.5 µm to 13 µm in the mid-infrared range (mid-IR). The system is based on a dual-band frequency domain optical parametric amplifier (FOPA) optically pumped by a Ti:Sapphire laser and amplifying 2 synchronized femtosecond pulses each with a widely tunable wavelength around 1.6 and 1.9 µm respectively. These amplified pulses are then combined in a GaSe crystal to produce the mid-IR few-cycle pulses by means of difference frequency generation (DFG). The architecture provides a passively stabilized carrier-envelope phase (CEP) whose fluctuations has been characterized to 370 mrad RMS.

Nonlinear microscopy and deep learning classification for mammary gland microenvironment studies.

Tumors, their microenvironment, and the mechanisms by which collagen morphology changes throughout cancer progression have recently been a topic of interest. Second harmonic generation (SHG) and polarization second harmonic (P-SHG) microscopy are label-free, hallmark methods that can highlight this alteration in the extracellular matrix (ECM). This article uses automated sample scanning SHG and P-SHG microscopy to investigate ECM deposition associated with tumors residing in the mammary gland. We show two different analysis approaches using the acquired images to distinguish collagen fibrillar orientation changes in the ECM. Lastly, we apply a supervised deep-learning model to classify naïve and tumor-bearing mammary gland SHG images. We benchmark the trained model using transfer learning with the well-known MobileNetV2 architecture. By fine-tuning the different parameters of these models, we show a trained deep-learning model that suits such a small dataset with 73% accuracy.

Ultrafast Resonant State Formation by the Coupling of Rydberg and Dark Autoionizing States.

Studying the dynamics of dark states is challenging due to their inability to undergo single-photon emission or absorption. This challenge is made even more difficult for dark autoionizing states owing to their ultrashort lifetime of a few femtoseconds. High-order harmonic spectroscopy recently appeared as a novel method to probe the ultrafast dynamics of a single atomic or molecular state. Here, we demonstrate the emergence of a new type of ultrafast resonance state as a manifestation of coupling between Rydberg and a dark autoionizing state dressed by a laser photon. Through high-order harmonic generation, this resonance results in extreme ultraviolet light emission that is more than one order of magnitude stronger than for the off-resonance case. The induced resonance can be leveraged to study the dynamics of a single dark autoionizing state and the transient changes in the dynamics of real states due to their overlap with the virtual laser-dressed states. In addition, the present results allow the generation of coherent ultrafast extreme ultraviolet light for advanced ultrafast science applications.

Second harmonic generation microscopy: a powerful tool for bio-imaging.

Second harmonic generation (SHG) microscopy is an important optical imaging technique in a variety of applications. This article describes the history and physical principles of SHG microscopy and its more advanced variants, as well as their strengths and weaknesses in biomedical applications. It also provides an overview of SHG and advanced SHG imaging in neuroscience and microtubule imaging and how these methods can aid in understanding microtubule formation, structuration, and involvement in neuronal function. Finally, we offer a perspective on the future of these methods and how technological advancements can help make SHG microscopy a more widely adopted imaging technique.

Femtosecond multimodal imaging with a laser-driven X-ray source.

Laser-plasma accelerators are compact linear accelerators based on the interaction of high-power lasers with plasma to form accelerating structures up to 1000 times smaller than standard radiofrequency cavities, and they come with an embedded X-ray source, namely betatron source, with unique properties: small source size and femtosecond pulse duration. A still unexplored possibility to exploit the betatron source comes from combining it with imaging methods able to encode multiple information like transmission and phase into a single-shot acquisition approach. In this work, we combine edge illumination-beam tracking (EI-BT) with a betatron X-ray source and present the demonstration of multimodal imaging (transmission, refraction, and scattering) with a compact light source down to the femtosecond timescale. The advantage of EI-BT is that it allows multimodal X-ray imaging technique, granting access to transmission, refraction and scattering signals from standard low-coherence laboratory X-ray sources in a single shot.

Analytic description and optimization of magneto-optical Kerr setups with photoelastic modulation.

Instruments based on the magneto-optical Kerr effect are routinely used to probe surface magnetic properties. These tools rely on the characterization of the polarization state of reflected light from the sample to collect information on its magnetization. Here, we present a theoretical optimization of common setups based on the magneto-optical Kerr effect. A detection scheme based on a simple analyzer and photodetector and one made from a polarizing beam splitter and balanced photodetectors are considered. The effect of including a photoelastic modulator (PEM) and a lock-in amplifier to detect the signal at harmonics of the modulating frequency is studied. Jones formalism is used to derive general expressions that link the intensity of the measured signal to the magneto-optical Fresnel reflection coefficients for any orientation of the polarizing optical components. Optimal configurations are then defined as those that allow measuring the Kerr rotation and ellipticity while minimizing nonmagnetic contributions from the diagonal Fresnel coefficients in order to improve the signal-to-noise ratio (SNR). The expressions show that with the PEM, setups based on polarizing beam splitters inherently offer a twofold higher signal than commonly used analyzers, and the experimental results confirm that the SNR is improved by more than 150%. Furthermore, we find that while all proposed detection schemes measure Kerr effects, only those with polarizing beam splitters allow measuring the Kerr rotation directly when no modulator is included. This accommodates, for instance, time-resolved measurements at relatively low laser pulse repetition rates. Ultrafast demagnetization measurements are presented as an example of such applications.

Phosphorylation of eIF4E in the stroma drives the production and spatial organisation of collagen type I in the mammary gland.

The extracellular matrix (ECM) plays critical roles in breast cancer development. Whether ECM composition is regulated by the phosphorylation of eIF4E on serine 209, an event required for tumorigenesis, has not been explored. Herein, we used proteomics and mouse modeling to investigate the impact of mutating serine 209 to alanine on eIF4E (i.e., S209A) on mammary gland (MG) ECM. The proteomic data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD028953. We discovered that S209A knock-in mice, expressing a non-phosphorylatable form of eIF4E, have less collagen-I deposition in native and tumor-bearing MGs, leading to altered tumor cell invasion. Additionally, phospho-eIF4E deficiency impacts collagen topology; fibers at the tumor-stroma boundary in phospho-eIF4E-deficient mice run parallel to the tumor edge but radiate outwards in wild-type mice. Finally, a phospho-eIF4E-deficient tumor microenvironment resists anti-PD-1 therapy-induced collagen deposition, correlating with an increased anti-tumor response to immunotherapy. Clinically, we showed that collagen-I and phospho-eIF4E are positively correlated in human breast cancer samples, and that stromal phospho-eIF4E expression is influenced by tumor proximity. Together, our work defines the importance of phosphorylation of eIF4E on S209 as a regulator of MG collagen architecture in the tumor microenvironment, thereby positioning phospho-eIF4E as a therapeutic target to augment response to therapy.

Post-Ionization Dynamics of the Polar Molecule OCS in Asymmetric Laser Fields.

We have investigated the dissociation mechanisms of the prototypical heavy polar molecule OCS into the two break-up channels of the dication, OCS2+ → O+ + CS+ and OC+ + S+, in phase-locked two-color intense laser fields. The branching ratio of the breaking of the C-O and C-S bonds followed a pronounced 2π-oscillation with a modulation depth of 11%, depending on the relative phase of the two-color laser fields. The fragment ejection direction of both break-up channels reflects the anisotropy of the tunneling ionization rate, following a 2π-periodicity, as well. The two dissociation pathways in the C-S bond breaking channel show different phase dependencies of the fragment ejection direction, which are assigned to post-ionization dynamics. These observations, resulting from the excitation with asymmetric two-color intense laser fields, supported by state-of-the-art theoretical simulations, reveal the importance of post-ionization population dynamics in addition to tunneling ionization in the molecular fragmentation processes, even for heavy polar molecules.

Spectral phase sensitivity of frequency resolved optical switching for broadband IR pulse characterization.

In this work, we demonstrate the sensitivity of the frequency-resolved optical switching (FROSt) technique to detect a small amount of spectral phase shift for the precise characterization of ultrashort laser pulses. We characterized fs pulses centered at 1.75 µm that are spectrally broadened up to 700 nm of bandwidth in a hollow-core fiber and subsequently compressed down to 2.3 optical cycle duration by propagation in the air at atmospheric pressure. By inserting thin fused silica windows of different thicknesses in the beam path, we accurately retrieve group delay dispersion (GDD) variations as small as 10 fs2. Such GDD variations correspond to a change of the pulse duration of only 0.2 fs for a Fourier transform limited 2-cycle pulse at 1.75 µm (i.e., 11.8 fs). The capability to measure such tiny temporal variations thus demonstrates that the FROSt technique has sufficient sensitivity to precisely characterize single-cycle pulses.

Electronic relaxation and dissociation dynamics in formaldehyde: pump wavelength dependence.

The effect of the incident UV pump wavelength on the subsequent excited state dynamics, electronic relaxation, and ultimate dissociation of formaldehyde is studied using first principles simulation and Coulomb explosion imaging (CEI) experiments. Transitions in a vibronic progression in the à ← X̃ absorption band are systematically prepared using a tunable UV source which generates pulses centered at 304, 314, 329, and 337 nm. We find, both via ab initio simulation and experimental results, that the rate of excited state decay and subsequent dissociation displays a prominent dependence on which vibronic transition in the absorption band is prepared by the pump. Our simulations predict that nonadiabatic transition rates and dissociation yields will increase by a factor of >100 as the pump wavelength is decreased from 337 to 304 nm. The experimental results and theoretical simulations are in broad agreement and both indicate that the dissociation rate plateaus rapidly after ≈2 ps following an ultrafast sub-ps rise.

Unsupervised hyperspectral stimulated Raman microscopy image enhancement: denoising and segmentation via one-shot deep learning.

Hyperspectral stimulated Raman scattering (SRS) microscopy is a label-free technique for biomedical and mineralogical imaging which can suffer from low signal-to-noise ratios. Here we demonstrate the use of an unsupervised deep learning neural network for rapid and automatic denoising of SRS images: UHRED (Unsupervised Hyperspectral Resolution Enhancement and Denoising). UHRED is capable of "one-shot" learning; only one hyperspectral image is needed, with no requirements for training on previously labelled datasets or images. Furthermore, by applying a k-means clustering algorithm to the processed data, we demonstrate automatic, unsupervised image segmentation, yielding, without prior knowledge of the sample, intuitive chemical species maps, as shown here for a lithium ore sample.

Polarization-independent pulse retrieval based on frequency resolved optical switching.

We demonstrate experimentally that the frequency resolved optical switching (FROSt) method is independent of the polarization direction of the pulse to be characterized. In this perspective, it is employed to characterize two or three co-propagating pulses linearly polarized in orthogonal directions, enabling to retrieve simultaneously their temporal intensity and phase profiles together with their group delay. This technique is also applied to track a simple nonlinear process involving different polarization states: type-I second harmonic generation (SHG). We are able to characterize the depleted fundamental pulse along with the generated second-harmonic pulse, thus demonstrating that the FROSt technique is a practical and powerful tool to observe nonlinear processes both in the temporal and spectral domains even if it involves different polarization states.

Time-resolved photoelectron imaging of complex resonances in molecular nitrogen.

We have used the FERMI free-electron laser to perform time-resolved photoelectron imaging experiments on a complex group of resonances near 15.38 eV in the absorption spectrum of molecular nitrogen, N2, under jet-cooled conditions. The new data complement and extend the earlier work of Fushitani et al. [Opt. Express 27, 19702-19711 (2019)], who recorded time-resolved photoelectron spectra for this same group of resonances. Time-dependent oscillations are observed in both the photoelectron yields and the photoelectron angular distributions, providing insight into the interactions among the resonant intermediate states. In addition, for most states, we observe an exponential decay of the photoelectron yield that depends on the ionic final state. This observation can be rationalized by the different lifetimes for the intermediate states contributing to a particular ionization channel. Although there are nine resonances within the group, we show that by detecting individual photoelectron final states and their angular dependence, we can identify and differentiate quantum pathways within this complex system.