Label-free drug interaction screening via Raman microscopy.

Development of a simple, label-free screening technique capable of precisely and directly sensing interaction-in-solution over a size range from small molecules to large proteins such as antibodies could offer an important tool for researchers and pharmaceutical companies in the field of drug development. In this work, we present a thermostable Raman interaction profiling (TRIP) technique that facilitates low-concentration and low-dose screening of binding between protein and ligand in physiologically relevant conditions. TRIP was applied to eight protein-ligand systems, and produced reproducible high-resolution Raman measurements, which were analyzed by principal component analysis. TRIP was able to resolve time-depending binding between 2,4-dinitrophenol and transthyretin, and analyze biologically relevant SARS-CoV-2 spike-antibody interactions. Mixtures of the spike receptor-binding domain with neutralizing, nonbinding, or binding but nonneutralizing antibodies revealed distinct and reproducible Raman signals. TRIP holds promise for the future developments of high-throughput drug screening and real-time binding measurements between protein and drug.

Label-free sensing of cells with fluorescence lifetime imaging: The quest for metabolic heterogeneity.

Molecular, morphological, and physiological heterogeneity is the inherent property of cells which governs differences in their response to external influence. Tumor cell metabolic heterogeneity is of a special interest due to its clinical relevance to tumor progression and therapeutic outcomes. Rapid, sensitive, and noninvasive assessment of metabolic heterogeneity of cells is a great demand for biomedical sciences. Fluorescence lifetime imaging (FLIM), which is an all-optical technique, is an emerging tool for sensing and quantifying cellular metabolism by measuring fluorescence decay parameters of endogenous fluorophores, such as NAD(P)H. To achieve accurate discrimination between metabolically diverse cellular subpopulations, appropriate approaches to FLIM data collection and analysis are needed. In this paper, the unique capability of FLIM to attain the overarching goal of discriminating metabolic heterogeneity is demonstrated. This has been achieved using an approach to data analysis based on the nonparametric analysis, which revealed a much better sensitivity to the presence of metabolically distinct subpopulations compared to more traditional approaches of FLIM measurements and analysis. The approach was further validated for imaging cultured cancer cells treated with chemotherapy. These results pave the way for accurate detection and quantification of cellular metabolic heterogeneity using FLIM, which will be valuable for assessing therapeutic vulnerabilities and predicting clinical outcomes.

Simultaneous Two- and Three-Photon Deep Imaging of Autofluorescence in Bacterial Communities.

The intrinsic fluorescence of bacterial samples has a proven potential for label-free bacterial characterization, monitoring bacterial metabolic functions, and as a mechanism for tracking the transport of relevant components through vesicles. The reduced scattering and axial confinement of the excitation offered by multiphoton imaging can be used to overcome some of the limitations of single-photon excitation (e.g., scattering and out-of-plane photobleaching) to the imaging of bacterial communities. In this work, we demonstrate in vivo multi-photon microscopy imaging of Streptomyces bacterial communities, based on the excitation of blue endogenous fluorophores, using an ultrafast Yb-fiber laser amplifier. Its parameters, such as the pulse energy, duration, wavelength, and repetition rate, enable in vivo multicolor imaging with a single source through the simultaneous two- and three-photon excitation of different fluorophores. Three-photon excitation at 1040 nm allows fluorophores with blue and green emission spectra to be addressed (and their corresponding ultraviolet and blue single-photon excitation wavelengths, respectively), and two-photon excitation at the same wavelength allows fluorophores with yellow, orange, or red emission spectra to be addressed (and their corresponding green, yellow, and orange single-photon excitation wavelengths). We demonstrate that three-photon excitation allows imaging over a depth range of more than 6 effective attenuation lengths to take place, corresponding to an 800 micrometer depth of imaging, in samples with a high density of fluorescent structures.

Spectral resolution enhancement for impulsive stimulated Brillouin spectroscopy by expanding pump beam geometry.

Brillouin microscopy has recently emerged as a powerful tool for mechanical property measurements in biomedical sensing and imaging applications. Impulsive stimulated Brillouin scattering (ISBS) microscopy has been proposed for faster and more accurate measurements, which do not rely on stable narrow-band lasers and thermally-drifting etalon-based spectrometers. However, the spectral resolution of ISBS-based signal has not been significantly explored. In this report, the ISBS spectral profile has been investigated as a function of the pump beam's spatial geometry, and novel methodologies have been developed for accurate spectral assessment. The ISBS linewidth was found to consistently decrease with increasing pump-beam diameter. These findings provide the means for improved spectral resolution measurements and pave the way to broader applications of ISBS microscopy.

Laser spectroscopic technique for direct identification of a single virus I: FASTER CARS.

From the famous 1918 H1N1 influenza to the present COVID-19 pandemic, the need for improved viral detection techniques is all too apparent. The aim of the present paper is to show that identification of individual virus particles in clinical sample materials quickly and reliably is near at hand. First of all, our team has developed techniques for identification of virions based on a modular atomic force microscopy (AFM). Furthermore, femtosecond adaptive spectroscopic techniques with enhanced resolution via coherent anti-Stokes Raman scattering (FASTER CARS) using tip-enhanced techniques markedly improves the sensitivity [M. O. Scully, et al, Proc. Natl. Acad. Sci. U.S.A. 99, 10994-11001 (2002)].

Non-Destructive Direct Pericarp Thickness Measurement of Sorghum Kernels Using Extended-Focus Optical Coherence Microscopy.

Non-destructive measurements of internal morphological structures in plant materials such as seeds are of high interest in agricultural research. The estimation of pericarp thickness is important to understand the grain quality and storage stability of seeds and can play a crucial role in improving crop yield. In this study, we demonstrate the applicability of fiber-based Bessel beam Fourier domain (FD) optical coherence microscopy (OCM) with a nearly constant high lateral resolution maintained at over ~400 µm for direct non-invasive measurement of the pericarp thickness of two different sorghum genotypes. Whereas measurements based on axial profiles need additional knowledge of the pericarp refractive index, en-face views allow for direct distance measurements. We directly determine pericarp thickness from lateral sections with a 3 µm resolution by taking the width of the signal corresponding to the pericarp at the 1/e threshold. These measurements enable differentiation of the two genotypes with 100% accuracy. We find that trading image resolution for acquisition speed and view size reduces the classification accuracy. Average pericarp thicknesses of 74 µm (thick phenotype) and 43 µm (thin phenotype) are obtained from high-resolution lateral sections, and are in good agreement with previously reported measurements of the same genotypes. Extracting the morphological features of plant seeds using Bessel beam FD-OCM is expected to provide valuable information to the food processing industry and plant breeding programs.

Ghost translation: an end-to-end ghost imaging approach based on the transformer network.

Artificial intelligence has recently been widely used in computational imaging. The deep neural network (DNN) improves the signal-to-noise ratio of the retrieved images, whose quality is otherwise corrupted due to the low sampling ratio or noisy environments. This work proposes a new computational imaging scheme based on the sequence transduction mechanism with the transformer network. The simulation database assists the network in achieving signal translation ability. The experimental single-pixel detector's signal will be 'translated' into a 2D image in an end-to-end manner. High-quality images with no background noise can be retrieved at a sampling ratio as low as 2%. The illumination patterns can be either well-designed speckle patterns for sub-Nyquist imaging or random speckle patterns. Moreover, our method is robust to noise interference. This translation mechanism opens a new direction for DNN-assisted ghost imaging and can be used in various computational imaging scenarios.

Synthesizing Five-Body Interaction in a Superconducting Quantum Circuit.

Synthesizing many-body interaction Hamiltonians is a central task in quantum simulation. However, it is challenging to synthesize Hamiltonians that have more than two spins in a single term. Here we synthesize m-body spin-exchange Hamiltonians with m up to 5 in a superconducting quantum circuit by simultaneously exciting multiple independent qubits with time-energy correlated photons generated from a qudit. The dynamic evolution of the m-body interaction is governed by the Rabi oscillation between two m-spin states, in which the states of each spin are different. We demonstrate the scalability of our approach by comparing the influence of noises on the three-, four- and five-body interaction and building a many-body Mach-Zehnder interferometer which potentially has a Heisenberg-limit sensitivity. This study paves a way for quantum simulation involving many-body interaction Hamiltonians such as lattice gauge theories in quantum circuits.

Sub-Rayleigh second-order correlation imaging using spatially distributive colored noise speckle patterns.

We present a novel method, to our knowledge, to synthesize non-trivial speckle patterns that can enable sub-Rayleigh second-order correlation imaging. The speckle patterns acquire a unique anti-correlation in the spatial intensity fluctuation by introducing the blue noise distribution on spatial Fourier power spectrum to the input light fields through amplitude modulation. Illuminating objects with the blue noise speckle patterns can lead to a sub-diffraction limit imaging system with a resolution more than three times higher than first-order imaging, which is comparable to the resolving power of ninth order correlation imaging with thermal light. Our method opens a new route towards non-trivial speckle pattern generation by tailoring amplitudes in spatial Fourier power spectrum of the input light fields and provides a versatile scheme for constructing sub-Rayleigh imaging and microscopy systems without invoking complicated higher-order correlations.

Unruh and Cherenkov Radiation from a Negative Frequency Perspective.

A ground-state atom uniformly accelerated through the Minkowski vacuum can become excited by emitting an Unruh-Minkowski photon. We show that from the perspective of an accelerated atom, the sign of the frequency of the Unruh-Minkowski photons can be positive or negative depending on the acceleration direction. The accelerated atom becomes excited by emitting an Unruh-Minkowski photon which has negative frequency in the atom's frame, and decays by emitting a positive-frequency photon. This leads to interesting effects. For example, the photon emitted by accelerated ground-state atom cannot be absorbed by another ground-state atom accelerating in the same direction, but it can be absorbed by an excited atom or a ground-state atom accelerated in the opposite direction. We also show that similar effects take place for Cherenkov radiation. Namely, a Cherenkov photon emitted by an atom cannot be absorbed by another ground-state atom moving with the same velocity, but can be absorbed by an excited atom or a ground-state atom moving in the opposite direction.

Two-level masers as heat-to-work converters.

Heat engines, which cyclically transform heat into work, are ubiquitous in technology. Lasers and masers may be viewed as heat engines that rely on population inversion or coherence in the active medium. Here we put forward an unconventional paradigm of a remarkably simple and robust electromagnetic heat-powered engine that bears basic differences to any known maser or laser: The proposed device makes use of only one Raman transition and does not rely on population inversion or coherence in its two-level working medium. Nor does it require any coherent driving. The engine can be powered by the ambient temperature difference between the sky and the ground surface. Its autonomous character and "free" power source make this engine conceptually and technologically enticing.

Quantum optics approach to radiation from atoms falling into a black hole.

We show that atoms falling into a black hole (BH) emit acceleration radiation which, under appropriate initial conditions, looks to a distant observer much like (but is different from) Hawking BH radiation. In particular, we find the entropy of the acceleration radiation via a simple laser-like analysis. We call this entropy horizon brightened acceleration radiation (HBAR) entropy to distinguish it from the BH entropy of Bekenstein and Hawking. This analysis also provides insight into the Einstein principle of equivalence between acceleration and gravity.

Insect flight velocity measurement with a CW near-IR Scheimpflug lidar system.

Flight velocity measurement is an important aspect of insect research that can aid insect identification and facilitate studies and monitoring of insect movements. We propose a novel scheme for the 1-D flight velocity measurement of insects, based on a near-IR Scheimpflug lidar system. We implement this new technique and apply it to study insects at the Salter Research Farm, Robertson County, Texas. The resolution property perpendicular to the probing direction of the Scheimpflug lidar system is explored and reveals the capability of retrieving the velocity component normal to the probing direction of insects passing through the field of view of our system. We observe a shift in wingbeat frequency, which indicates the presence of new insect species during the multi-day measurement. The study on 1-D flight velocity reveals a net directional movement of insects, providing supportive evidence of the arrival of a new species.

Light and corona: guided-wave readout for coronavirus spike protein-host-receptor binding.

We show that waveguide sensors can enable a quantitative characterization of coronavirus spike glycoprotein-host-receptor binding-the process whereby coronaviruses enter human cells, causing disease. We demonstrate that such sensors can help quantify and eventually understand kinetic and thermodynamic properties of viruses that control their affinity to targeted cells, which is known to significantly vary in the course of virus evolution, e.g., from SARS-CoV to SARS-CoV-2, making the development of virus-specific drugs and vaccine difficult. With the binding rate constants and thermodynamic parameters as suggested by the latest SARS-CoV-2 research, optical sensors of SARS-CoV-2 spike protein-receptor binding may be within sight.

Simultaneous Excitation of Two Noninteracting Atoms with Time-Frequency Correlated Photon Pairs in a Superconducting Circuit.

We report the first observation of simultaneous excitation of two noninteracting atoms by a pair of time-frequency correlated photons in a superconducting circuit. The strong coupling regime of this process enables the synthesis of a three-body interaction Hamiltonian, which allows the generation of the tripartite Greenberger-Horne-Zeilinger state in a single step with a fidelity as high as 0.95. We further demonstrate the inhibition of the simultaneous two-atom excitation by continuously measuring whether the first photon is emitted. This work provides a new route in synthesizing many-body interaction Hamiltonian and coherent control of entanglement.

In vivo diagnostics of early abiotic plant stress response via Raman spectroscopy.

Development of a phenotyping platform capable of noninvasive biochemical sensing could offer researchers, breeders, and producers a tool for precise response detection. In particular, the ability to measure plant stress in vivo responses is becoming increasingly important. In this work, a Raman spectroscopic technique is developed for high-throughput stress phenotyping of plants. We show the early (within 48 h) in vivo detection of plant stress responses. Coleus (Plectranthus scutellarioides) plants were subjected to four common abiotic stress conditions individually: high soil salinity, drought, chilling exposure, and light saturation. Plants were examined poststress induction in vivo, and changes in the concentration levels of the reactive oxygen-scavenging pigments were observed by Raman microscopic and remote spectroscopic systems. The molecular concentration changes were further validated by commonly accepted chemical extraction (destructive) methods. Raman spectroscopy also allows simultaneous interrogation of various pigments in plants. For example, we found a unique negative correlation in concentration levels of anthocyanins and carotenoids, which clearly indicates that plant stress response is fine-tuned to protect against stress-induced damages. This precision spectroscopic technique holds promise for the future development of high-throughput screening for plant phenotyping and the quantification of biologically or commercially relevant molecules, such as antioxidants and pigments.

Enhanced coupling of light into a turbid medium through microscopic interface engineering.

There are many optical detection and sensing methods used today that provide powerful ways to diagnose, characterize, and study materials. For example, the measurement of spontaneous Raman scattering allows for remote detection and identification of chemicals. Many other optical techniques provide unique solutions to learn about biological, chemical, and even structural systems. However, when these systems exist in a highly scattering or turbid medium, the optical scattering effects reduce the effectiveness of these methods. In this article, we demonstrate a method to engineer the geometry of the optical interface of a turbid medium, thereby drastically enhancing the coupling efficiency of light into the material. This enhanced optical coupling means that light incident on the material will penetrate deeper into (and through) the medium. It also means that light thus injected into the material will have an enhanced interaction time with particles contained within the material. These results show that, by using the multiple scattering of light in a turbid medium, enhanced light-matter interaction can be achieved; this has a direct impact on spectroscopic methods such as Raman scattering and fluorescence detection in highly scattering regimes. Furthermore, the enhanced penetration depth achieved by this method will directly impact optical techniques that have previously been limited by the inability to deposit sufficient amounts of optical energy below or through highly scattering layers.

Tip-Enhanced Raman Imaging of Single-Stranded DNA with Single Base Resolution.

Tip-enhanced Raman scattering (TERS) is a promising optical and analytical technique for chemical imaging and sensing at single molecule resolution. In particular, TERS signals generated by a gap-mode configuration where a silver tip is coupled with a gold substrate can resolve a single-stranded DNA (ssDNA) molecule with a spatial resolution below 1 nm. To demonstrate the proof of subnanometer resolution, we show direct nucleic acid sequencing using TERS of a phage ssDNA (M13mp18). M13mp18 provides a known sequence and, through our deposition strategy, can be stretched (uncoiled) and attached to the substrate by its phosphate groups, while exposing its nucleobases to the tip. After deposition, we scan the silver tip along the ssDNA and collect TERS signals with a step of 0.5 nm, comparable to the bond length between two adjacent DNA bases. By demonstrating the real-time profiling of a ssDNA configuration and furthermore, with unique TERS signals of monomeric units of other biopolymers, we anticipate that this technique can be extended to the high-resolution imaging of various nanostructures as well as the direct sequencing of other important biopolymers including RNA, polysaccharides, and polypeptides.

Enhanced signals from chiral molecules via molecular coherence.

One of the most widely used chiroptical spectroscopic methods for studying chiral molecules is Raman optical activity; however, the chiral Raman optical activity signal is extremely weak. Here, we theoretically examine enhanced chiral signals in a system with strongly prepared molecular coherence. We show that the enhanced chiral signal due to strong molecular coherence is up to four orders of magnitude higher than that of the spontaneous Raman optical activity. We discuss several advantages of studying the heterodyned signal obtained by combining the anti-Stokes signal with a local oscillator. The heterodyning allows direct measurement of the ratio of the chiral and achiral parameters. Taking advantage of the molecular coherence and heterodyne detection, the coherent anti-Stokes Raman scattering technique opens up a new potential application for investigation of biomolecular chirality.

Highly efficient tunable picosecond deep ultraviolet laser system for Raman spectroscopy.

We present a narrowband laser system tunable from 219 to 236 nm for deep ultraviolet (DUV) Raman spectroscopy. The demonstrated laser system produces 6.7 ps nearly transform-limited pulses with energy up to 0.36 µJ at 100 kHz repetition rate. The system consists of a two-stage optical parametric amplifier (OPA) of a narrowband continuous wave diode laser and subsequent frequency conversion to the DUV radiation. We achieve more than 300 mW in the signal wave using ${{\rm LiB}_3}{{\rm O}_5}$LiB3O5 (LBO) and ${{\rm BaB}_2}{{\rm O}_4}$BaB2O4 (BBO) crystals, with the total 2.7 W pump after the two-stage OPA. We reach 12% conversion efficiency of the OPA signal wave into the DUV radiation using type-I phase matching in the BBO crystal. Finally, we demonstrate the applicability of the system for DUV Raman spectroscopy by collecting a high dynamic range, high spectral resolution spontaneous Raman spectrum of air.