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Spontaneous Raman microscopy is well-known for its remarkable chemical contrast yet suffers from slow acquisition speeds. Recently, the compressive Raman microspectroscopy framework has shown that a significant speed advantage is brought by leveraging shot-noise-limited detection using a single-photon avalanche diode (SPAD). However, current imaging speeds of compressive Raman architectures are fundamentally limited by SPAD sensitivity and dead time. Here, we demonstrate an efficient and scalable compressive Raman parallelization scheme based on SPAD arrays. We show that parallelization using line excitation, instead of spatial multiplexing, allows to reach effective pixel dwell times ( τ pdt ) of 0.8 µs. Such fast speed represents over one order-of-magnitude speed-up over previous demonstrations. This effective parallelization not only allows for demonstrating unprecedented chemical imaging speeds using the otherwise weak spontaneous Raman effect but also paves the way for true video-rate inexpensive molecular microspectroscopy.

Transition metal oxides are state-of-the-art materials for catalysing the oxygen evolution reaction (OER), whose slow kinetics currently limit the efficiency of water electrolysis. However, microscale physicochemical heterogeneity between particles, dynamic reactions both in the bulk and at the surface, and an interplay between particle reactivity and electrolyte makes probing the OER challenging. Here, we overcome these limitations by applying state-of-the-art compressive Raman imaging to uncover concurrent bias-dependent pathways for the OER in a dense, crystalline electrocatalyst, α-Li 2 IrO 3 . By spatially and temporally tracking changes in stretching modes we follow catalytic activation and charge accumulation following ion exchange under various electrolytes and cycling conditions, comparing our observations with other crystalline catalysts (IrO 2 , LiCoO 2 ). We demonstrate that at low overpotentials the reaction between water and the oxidized catalyst surface is compensated by bulk ion exchange, as usually only found for amorphous, electrolyte permeable, catalysts. At high overpotentials the charge is compensated by surface redox active sites, as in other crystalline catalysts such as IrO 2 . Hence, our work reveals charge compensation can extend beyond the surface in crystalline catalysts. More generally, the results highlight the power of compressive Raman imaging for chemically specific tracking of microscale reaction dynamics in catalysts, battery materials, or memristors., (© 2024. The Author(s).)

The intricate nature of the brain necessitates the application of advanced probing techniques to comprehensively study and understand its working mechanisms. Neurophotonics offers minimally invasive methods to probe the brain using optics at cellular and even molecular levels. However, multiple challenges persist, especially concerning imaging depth, field of view, speed, and biocompatibility. A major hindrance to solving these challenges in optics is the scattering nature of the brain. This perspective highlights the potential of complex media optics, a specialized area of study focused on light propagation in materials with intricate heterogeneous optical properties, in advancing and improving neuronal readouts for structural imaging and optical recordings of neuronal activity. Key strategies include wavefront shaping techniques and computational imaging and sensing techniques that exploit scattering properties for enhanced performance. We discuss the potential merger of the two fields as well as potential challenges and perspectives toward longer term in vivo applications., (© 2024 The Authors.)

Imaging at depth in opaque materials has long been a challenge. Recently, wavefront shaping has enabled notable advance for deep imaging. Nevertheless, most noninvasive wavefront-shaping methods require cameras, lack the sensitivity for deep imaging under weak optical signals, or can only focus on a single "guidestar." Here, we retrieve the transmission matrix (TM) noninvasively using two-photon fluorescence exploiting a single-pixel detection combined with a computational framework, allowing to achieve single-target focus on multiple guidestars spread beyond the memory effect range. In addition, if we assume that memory effect correlations exist in the TM, we are able to substantially reduce the number of measurements needed.

Computational methods have been established as cornerstones in optical imaging and holography in recent years. Every year, the dependence of optical imaging and holography on computational methods is increasing significantly to the extent that optical methods and components are being completely and efficiently replaced with computational methods at low cost. This roadmap reviews the current scenario in four major areas namely incoherent digital holography, quantitative phase imaging, imaging through scattering layers, and super-resolution imaging. In addition to registering the perspectives of the modern-day architects of the above research areas, the roadmap also reports some of the latest studies on the topic. Computational codes and pseudocodes are presented for computational methods in a plug-and-play fashion for readers to not only read and understand but also practice the latest algorithms with their data. We believe that this roadmap will be a valuable tool for analyzing the current trends in computational methods to predict and prepare the future of computational methods in optical imaging and holography., Supplementary Information: The online version contains supplementary material available at 10.1007/s00340-024-08280-3., Competing Interests: Conflict of interestThe authors declare no competing interests., (© The Author(s) 2024.)

Understanding (de)lithiation heterogeneities in battery materials is key to ensure optimal electrochemical performance. However, this remains challenging due to the three-dimensional morphology of electrode particles, the involvement of both solid- and liquid-phase reactants and a range of relevant timescales (seconds to hours). Here we overcome this problem and demonstrate the use of confocal microscopy for the simultaneous three-dimensional operando measurement of lithium-ion dynamics in individual agglomerate particles, and the electrolyte in batteries. We examine two technologically important cathode materials: Li x CoO 2 and Li x Ni 0.8 Mn 0.1 Co 0.1 O 2 . The surface-to-core transport velocity of Li-phase fronts and volume changes are captured as a function of cycling rate. Additionally, we visualize heterogeneities in the bulk and at agglomerate surfaces during cycling, and image microscopic liquid electrolyte concentration gradients. We discover that surface-limited reactions and intra-agglomerate competing rates control (de)lithiation and structural heterogeneities in agglomerate-based electrodes. Importantly, the conditions under which optical imaging can be performed inside the complex environments of battery electrodes are outlined., (© 2023. © The Author(s), under exclusive license to Springer Nature Limited.)

Photobatteries, batteries with a light-sensitive electrode, have recently been proposed as a way of simultaneously capturing and storing solar energy in a single device. Despite reports of photocharging with multiple different electrode materials, the overall mechanism of operation remains poorly understood. Here, we use operando optical reflection microscopy to investigate light-induced charging in Li x V 2 O 5 electrodes. We image the electrode, at the single-particle level, under three conditions: (a) with a closed circuit and light but no electronic power source (photocharging), (b) during galvanostatic cycling with light (photoenhanced), and (c) with heat but no light (thermal). We demonstrate that light can indeed drive lithiation changes in Li x V 2 O 5 while maintaining charge neutrality, possibly via a combination of faradaic and nonfaradaic effects taking place in individual particles. Our results provide an addition to the photobattery mechanistic model highlighting that both intercalation-based charging and lithium concentration polarization effects contribute to the increased photocharging capacity.

Unlike the interface between two immiscible electrolyte solutions (ITIES) formed between water and polar solvents, molecular understanding of the liquid-liquid interface formed for aqueous biphasic systems (ABSs) is relatively limited and mostly relies on surface tension measurements and thermodynamic models. Here, high-resolution Raman imaging is used to provide spatial and chemical resolution of the interface of lithium chloride - lithium bis(trifluoromethanesulfonyl)imide - water (LiCl-LiTFSI-water) and HCl-LiTFSI-water, prototypical salt-salt ABSs found in a range of electrochemical applications. The concentration profiles of both TFSI anions and water are found to be sigmoidal thus not showing any signs of a positive adsorption for both salts and solvent. More striking, however, is the length at which the concentration profiles extend, ranging from 11 to 2 µm with increasing concentrations, compared to a few nanometers for ITIES. We thus reveal that unlike ITIES, salt-salt ABSs do not have a molecularly sharp interface but rather form an interphase with a gradual change of environment from one phase to the other. This knowledge represents a major stepping-stone in the understanding of aqueous interfaces, key for mastering ion or electron transfer dynamics in a wide range of biological and technological settings including novel battery technologies such as membraneless redox flow and dual-ion batteries.

Three-photon (3P) microscopy is getting traction due to its superior performance in deep tissues. Yet, aberrations and light scattering still pose one of the main limitations in the attainable depth ranges for high-resolution imaging. Here, we show scattering correcting wavefront shaping with a simple continuous optimization algorithm, guided by the integrated 3P fluorescence signal. We demonstrate focusing and imaging behind scattering layers and investigate convergence trajectories for different sample geometries and feedback non-linearities. Furthermore, we show imaging through a mouse skull and demonstrate a novel, to the best of our knowledge, fast phase estimation scheme that substantially increases the speed at which the optimal correction can be found.

Raman scattering is a chemically selective probing mechanism with diverse applications in industry and clinical settings. Yet, most samples are optically opaque limiting the applicability of Raman probing at depth. Here, we demonstrate chemically selective energy deposition behind a scattering medium by combining prior information on the chemical's spectrum with the measurement of a spectrally resolved Raman speckle as a feedback mechanism for wavefront shaping. We demonstrate unprecedented sixfold signal enhancement in an epi-geometry, realizing targeted energy deposition and focusing on individual Raman active particles.

Non-invasive optical imaging techniques are essential diagnostic tools in many fields. Although various recent methods have been proposed to utilize and control light in multiple scattering media, non-invasive optical imaging through and inside scattering layers across a large field of view remains elusive due to the physical limits set by the optical memory effect, especially without wavefront shaping techniques. Here, we demonstrate an approach that enables non-invasive fluorescence imaging behind scattering layers with field-of-views extending well beyond the optical memory effect. The method consists in demixing the speckle patterns emitted by a fluorescent object under variable unknown random illumination, using matrix factorization and a novel fingerprint-based reconstruction. Experimental validation shows the efficiency and robustness of the method with various fluorescent samples, covering a field of view up to three times the optical memory effect range. Our non-invasive imaging technique is simple, neither requires a spatial light modulator nor a guide star, and can be generalized to a wide range of incoherent contrast mechanisms and illumination schemes., (© 2022. The Author(s).)

Propagation of an ultrashort pulse of light through strongly scattering media generates an intricate spatio-spectral speckle that can be described by means of the multi-spectral transmission matrix (MSTM). In conjunction with a spatial light modulator, the MSTM enables the manipulation of the pulse leaving the medium; in particular focusing it at any desired spatial position and/or time. Here, we demonstrate how to engineer the point-spread-function of the focused beam both spatially and spectrally, from the measured MSTM. It consists of numerically filtering the spatial content at each wavelength of the matrix prior to focusing. We experimentally report on the versatility of the technique through several examples, in particular as an alternative to simultaneous spatial and temporal focusing, with potential applications in multiphoton microscopy.

Hydrophobic oil droplets, particles, and air bubbles can be dispersed in water as kinetically stabilized dispersions. It has been established since the 19th century that such objects harbor a negative electrostatic potential roughly twice larger than the thermal energy. The source of this charge continues to be one of the core observations in relation to hydrophobicity, and its molecular explanation is still debated. What is clear though is that the stabilizing interaction in these systems is understood in terms of electrostatic repulsion via Derjaguin, Landau, Verwey, and Overbeek theory. Recent work [A. P. Carpenter et al., Proc. Natl. Acad. Sci. U. S. A. 116, 9214 (2019)] has added another element into the discussion, reporting the creation of bare near-zero charged droplets of oil in neat water that are stable for several days. Key to the creation of the droplets is a rigorous glassware cleaning procedure. Here, we investigate these conclusions and show that the cleaning procedure of glassware has no influence on the electrophoretic mobility of the droplets and that oil droplets with near-zero charge are unstable. We provide an alternative possible explanation for the observations involving glass surface chemistry.

The properties of water at an electrified graphene electrode are studied via classical molecular dynamics simulations with a constant potential approach. We show that the value of the applied electrode potential has dramatic effects on the structure and dynamics of interfacial water molecules. While a positive potential slows down the reorientational and translational dynamics of water, an increasing negative potential first accelerates the interfacial water dynamics before a deceleration at very large magnitude potential values. Further, our spectroscopic calculations indicate that the water rearrangements induced by electrified interfaces can be probed experimentally. In particular, the calculated water vibrational sum-frequency generation (SFG) spectra show that SFG specifically reports on the first two water layers at 0 V but that at larger magnitude applied potentials the resulting static field induces long-range contributions to the spectrum. Electrified graphene interfaces provide promising paradigm systems for comprehending both short- and long-range neighboring aqueous system impacts.

Advances in the research of intermolecular and surface interactions result from the development of new and improved measurement techniques and combinations of existing techniques. Here, we present a new miniature version of the surface forces apparatus-the μSFA-that has been designed for ease of use and multimodal capabilities with the retention of the capabilities of other SFA models including accurate measurements of the surface separation distance and physical characterization of dynamic and static physical forces (i.e., normal, shear, and friction) and interactions (e.g., van der Waals, electrostatic, hydrophobic, steric, and biospecific). The small physical size of the μSFA, compared to previous SFA models, makes it portable and suitable for integration into commercially available optical and fluorescence light microscopes, as demonstrated here. The large optical path entry and exit ports make it ideal for concurrent force measurements and spectroscopy studies. Examples of the use of the μSFA in combination with surface plasmon resonance (SPR) and Raman spectroscopy measurements are presented. Because of the short working distance constraints associated with Raman spectroscopy, an interferometric technique was developed and applied to calculate the intersurface separation distance based on Newton's rings. The introduction of the μSFA will mark a transition in SFA usage from primarily physical characterization to concurrent physical characterization with in situ chemical and biological characterization to study interfacial phenomena, including (but not limited to) molecular adsorption, fluid flow dynamics, the determination of surface species and morphology, and (bio)molecular binding kinetics.

Modern interfacial science is increasingly multidisciplinary. Unique insight into interfacial interactions requires new multimodal techniques for interrogating surfaces with simultaneous complementary physical and chemical measurements. Here, we describe the design and testing of a microscope that incorporates a miniature surface forces apparatus (μSFA) in sphere vs flat geometry for force-distance measurements, while simultaneously acquiring Raman spectra of the confined zone. The simple optical setup isolates independent optical paths for (i) the illumination and imaging of Newton's rings and (ii) Raman scattering excitation and efficient signal collection. We benchmark the methodology by examining Teflon thin films in asymmetric (Teflon-water-glass) and symmetric (Teflon-water-Teflon) configurations. Water is observed near the Teflon-glass interface with nanometer-scale sensitivity in both the distance and Raman signals. We perform chemically resolved, label-free imaging of confined contact regions between Teflon and glass surfaces immersed in water. Remarkably, we estimate that the combined approach enables vibrational spectroscopy with single water monolayer sensitivity within minutes. Altogether, the Raman-μSFA allows exploration of molecular confinement between surfaces with chemical selectivity and correlation with interaction forces.

Wavefront shaping is a powerful method to refocus light through a scattering medium. Its application to large spectral bandwidths or multiple wavelengths refocusing for nonlinear bio-imaging in-depth is however limited by spectral decorrelations. In this work, we demonstrate ways to access a large spectral memory of a refocus in thin scattering media and thick forward-scattering biological tissues. First, we show that the accessible spectral bandwidth through a scattering medium involves an axial spatio-spectral coupling, which can be minimized when working in a confocal geometry. Second, we show that this bandwidth can be further enlarged when working in a broadband excitation regime. These results open important prospects for multispectral nonlinear imaging through scattering media.

We experimentally implement a compressive Raman technology (CRT) that incorporates chemometric analysis directly into the spectrometer hardware by means of a digital micromirror device (DMD). The DMD is a programmable optical filter on which optimized binary filters are displayed. The latter are generated with an algorithm based on the Cramer-Rao lower bound. We compared the developed CRT microspectrometer with two conventional state-of-the-art Raman hyperspectral imaging systems on samples mimicking microcalcifications relevant for breast cancer diagnosis. The CRT limit of detection significantly improves, when compared to the CCD based system, and CRT ultimately allows 100× and 10× faster acquisition speeds than the CCD- and EMCCD-based systems, respectively.

Over recent decades, lipid membranes have become standard models for examining the biophysics and biochemistry of cell membranes. Interrogation of lipid domains within biomembranes is generally done with fluorescence microscopy via exogenous chemical probes. However, fluorophores have limited partitioning tunability, with the majority segregating into the liquid-disordered phase, and fluorescence only strictly reports on the small percentage of tagged lipids. We present simple, label-free imaging of domain formation in lipid monolayers, with chemical selectivity in unraveling lipid and cholesterol composition in different domain types. Exploiting conventional vibrational contrast in spontaneous Raman imaging, combined with chemometrics analysis, allows for examination of ternary systems containing saturated lipids, unsaturated lipids, and cholesterol. We confirm features commonly observed by fluorescence microscopy and provide a quantitative thermodynamic analysis of cholesterol distribution at the single-monolayer level.

The precision of proportion estimation with binary filtering of a Raman spectrum mixture is analyzed when the number of binary filters is equal to the number of present species and when the measurements are corrupted with Poisson photon noise. It is shown that the Cramer-Rao bound provides a useful methodology to analyze the performance of such an approach, in particular when the binary filters are orthogonal. It is demonstrated that a simple linear mean square error estimation method is efficient (i.e., has a variance equal to the Cramer-Rao bound). Evolutions of the Cramer-Rao bound are analyzed when the measuring times are optimized or when the considered proportion for binary filter synthesis is not optimized. Two strategies for the appropriate choice of this considered proportion are also analyzed for the binary filter synthesis.

The control and use of light polarization in optical sciences and engineering are widespread. Despite remarkable developments in polarization-resolved imaging for life sciences, their transposition to strongly scattering media is currently not possible, because of the inherent depolarization effects arising from multiple scattering. We show an unprecedented phenomenon that opens new possibilities for polarization-resolved microscopy in strongly scattering media: polarization recovery via broadband wavefront shaping. We demonstrate focusing and recovery of the original injected polarization state without using any polarizing optics at the detection. To enable molecular-level structural imaging, an arbitrary rotation of the input polarization does not degrade the quality of the focus. We further exploit the robustness of polarization recovery for structural imaging of biological tissues through scattering media. We retrieve molecular-level organization information of collagen fibers by polarization-resolved second harmonic generation, a topic of wide interest for diagnosis in biomedical optics. Ultimately, the observation of this new phenomenon paves the way for extending current polarization-based methods to strongly scattering environments.

We report a simple add-on for broadband stimulated Raman scattering (SRS) microscopes to enable fast and programmable spectroscopy acquisition. It comprises a conventional dispersive spectrometer layout incorporating a fast digital micromirror device (DMD). The approach is validated by acquiring SRS spectra of standard chemicals. We demonstrate a DMD's advantage in broadband SRS by showing higher signal-to-noise ratio using a multiplexed Hadamard spectral basis and compressive sensing detection. Our results apply to a variety of frequency-domain pump-probe spectroscopy.

Polarization resolved nonlinear microscopy (PRNM) is a powerful technique to gain microscopic structural information in biological media. However, deep imaging in a variety of biological specimens is hindered by light scattering phenomena, which not only degrades the image quality but also affects the polarization state purity. In order to quantify this phenomenon and give a framework for polarization resolved microscopy in thick scattering tissues, we develop a characterization methodology based on four wave mixing (FWM) process. More specifically, we take advantage of two unique features of FWM, meaning its ability to produce an intrinsic in-depth local coherent source and its capacity to quantify the presence of light depolarization in isotropic regions inside a sample. By exploring diverse experimental layouts in phantoms with different scattering properties, we study systematically the influence of scattering on the nonlinear excitation and emission processes. The results show that depolarization mechanisms for the nonlinearly generated photons are highly dependent on the scattering center size, the geometry used (epi/forward) and, most importantly, on the thickness of the sample. We show that the use of an un-analyzed detection makes the polarization-dependence read-out highly robust to scattering effects, even in regimes where imaging might be degraded. The effects are illustrated in polarization resolved imaging of myelin lipid organization in mouse spinal cords.

Specific ion effects can influence many processes in aqueous solutions: protein folding, enzyme activity, self-assembly, and interface stabilization. Ionic amphiphiles are known to stabilize the oil/water interface, presumably by dipping their hydrophobic tails into the oil phase while sticking their hydrophilic head groups in water. However, we find that anionic and cationic amphiphiles adopt strikingly different structures at liquid hydrophobic/water interfaces, linked to the different specific interactions between water and the amphiphile head groups, both at the interface and in the bulk. Vibrational sum frequency scattering measurements show that dodecylsulfate (DS(-)) ions do not detectably perturb the oil phase while dodecyltrimethylammonium (DTA(+)) ions do. Raman solvation shell spectroscopy and second harmonic scattering (SHS) show that the respective hydration-shells and the interfacial water structure are also very different. Our work suggests that specific interactions with water play a key role in driving the anionic head group toward the water phase and the cationic head group toward the oil phase, thus also implying a quite different surface stabilization mechanism.

Ultralow densities of crystalline nanospheres have been detected in amorphous polymer microspheres by utilizing the unique sensitivity of second-order nonlinear optical techniques to anisotropy. Vibrational sum frequency scattering (SFS) and X-ray diffraction (XRD) are used to quantify the crystallinity of amorphous poly(d,l-lactic acid) microspheres. While XRD does not display any crystallinity for the microspheres, SFS spectra and patterns are reminiscent of a heterogeneous microsphere that contains small crystalline domains. Nonlinear light scattering theory was used to model the data, and an average domain radius of 147 ± 65 nm was obtained. The degree of crystallinity (0.2%) was estimated by comparing XRD and SFS data obtained from the amorphous microspheres to similar data obtained from crystalline microspheres. We estimate a detection limit of 0.002% for SFS.

We present a comparison between sum frequency scattering (SFS) and reflection mode sum frequency generation (R-SFG). We have used scattering theory to describe both scattering experiments as well as reflection mode experiments. The interfacial vibrational spectrum of nanoscopic oil droplets dispersed in water was probed with SFS as well as with R-SFG. Spectra recorded in phase-matched R-SFG mode and spectra recorded with SFS from the same sample are different, which shows that different interfaces are measured. Scattering spectra at different scattering angles agree with nonlinear light scattering theory. We further present experiments with polymer films aimed at quantifying the comparative strength of R-SFG and SFS experiments.

We established the charge and structure of the oil/water interface by combining ζ-potential measurements, sum frequency scattering (SFS) and molecular dynamics simulations. The SFS experiments show that the orientation of water molecules can be followed on the oil droplet/water interface. The average water orientation on a neat oil droplet/water interface is the same as the water orientation on a negatively charged interface. pH dependent experiments show, however, that there is no sign of selective adsorption of hydroxide ions. Molecular dynamics simulations, both with and without intermolecular charge transfer, show that the balance of accepting and donating hydrogen bonds is broken in the interfacial layer, leading to surface charging. This can account for the negative surface charge that is found in experiments.

We present sum frequency scattering spectra on kinetically stabilized emulsions consisting of nanoscopic oil droplets in water, stabilized with sodium dodecyl sulfate (SDS). We have measured the interfacial structure of the alkyl chains of the surfactant molecules, the alkyl chain of the oil molecules, the weakly dispersive D(2)O response, and the interference between SDS and the oil. We find a big difference in chain conformation: SDS has many chain defects, whereas the oil has very few. Our spectra are interpreted to originate from a surface structure with oil molecules predominantly oriented parallel with respect to the plane of the interface. The SDS headgroup is surrounded by water molecules. The SDS alkyl tail is in a disordered state and partially in contact with water. Such a conformation of surfactant occupies a surface area of several hundreds of squared angstroms.

Surfactants such as sodium dodecylsulfate (SDS) can reduce the interfacial tension between bulk water and bulk n-hexadecane by 42 mN/m. Although reduction of interfacial tension should also take place on the interface of nanoscopic oil droplets in water, vibrational sum frequency scattering experiments indicate otherwise. In these measurements we have directly measured the adsorption of SDS onto hexadecane oil droplets with an average radius of 83 nm. We find that the interfacial density of adsorbed SDS is at least 1 order of magnitude lower than that at a corresponding planar interface. The derived maximum decrease in interfacial tension is only 5 mN/m.

We present sum-frequency-scattering experiments on colloidal dispersions with various concentrations and in different scattering geometries. At small scattering angles, large fluctuations are observed in the intensity of the scattered sum-frequency photons. By considering the angular dependence of the signal, the particle concentration dependence, and the surface vibrational spectra of the particle, we have determined that the fluctuations are caused by scattering from clusters of particles. We further demonstrate that dynamic nonlinear light scattering may be used to measure the size of the correlated particle clusters.

Many processes in chemistry and physics rely on the structure, growth or change of material buried in solids. The impenetrable surrounding medium often prohibits the study of such material in situ. Nonlinear light scattering can be used to observe the internal structure of a crystalline state embedded inside another solid state. Vibrational sum frequency scattering patterns of polymer microspheres, consisting of both amorphous and crystalline material, reveal the size of the buried microstructure and the optical components of the second-order susceptibility of the material. The vibrational spectra reveal the molecular structure.