Simulation of optomechanical interaction of levitated nanoparticle with photonic crystal micro cavity.

We propose and analyze theoretically a promising design of an optical trap for vacuum levitation of nanoparticles based on a one-dimensional (1D) silicon photonic crystal cavity (PhC). The considered cavity has a quadratically modulated width of the silicon wave guiding structure, leading to a calculated cavity quality factor of 8 × 105. An effective mode volume of approximately 0.16 µm3 having the optical field strongly confined outside the silicon structure enables optical confinement on nanoparticle in all three dimensions. The optical forces and particle-cavity optomechanical coupling are comprehensively analyzed for two sizes of silica nanoparticles (100 nm and 150 nm in diameter) and various mode detunings. The value of trapping stiffnesses in the microcavity is predicted to be 5 order of magnitudes higher than that reached for optimized optical tweezers, moreover the linear single photon coupling rate can reach MHz level which is 6 order magnitude larger than previously reported values for common bulk cavities. The theoretical results support optimistic prospects towards a compact chip for optical levitation in vacuum and cooling of translational mechanical degrees of motion for the silica nanoparticle of a diameter of 100 nm.

Physiological and transcriptome profiling of Chlorella sorokiniana: A study on azo dye wastewater decolorization.

Over decades, synthetic dyes have become increasingly dominated by azo dyes posing a significant environmental risk due to their toxicity. Microalgae-based systems may offer an alternative for treatment of azo dye effluents to conventional physical-chemical methods. Here, microalgae were tested to decolorize industrial azo dye wastewater (ADW). Chlorella sorokiniana showed the highest decolorization efficiency in a preliminary screening test. Subsequently, the optimization of the experimental design resulted in 70% decolorization in a photobioreactor. Tolerance of this strain was evidenced using multiple approaches (growth and chlorophyll content assays, scanning electron microscopy (SEM), and antioxidant level measurements). Raman microspectroscopy was employed for the quantification of ADW-specific compounds accumulated by the microalgal biomass. Finally, RNA-seq revealed the transcriptome profile of C. sorokiniana exposed to ADW for 72 h. Activated DNA repair and primary metabolism provided sufficient energy for microalgal growth to overcome the adverse toxic conditions. Furthermore, several transporter genes, oxidoreductases-, and glycosyltransferases-encoding genes were upregulated to effectively sequestrate and detoxify the ADW. This work demonstrates the potential utilization of C. sorokiniana as a tolerant strain for industrial wastewater treatment, emphasizing the regulation of its molecular mechanisms to cope with unfavorable growth conditions.

Synchronization of spin-driven limit cycle oscillators optically levitated in vacuum.

We explore, experimentally and theoretically, the emergence of coherent coupled oscillations and synchronization between a pair of non-Hermitian, stochastic, opto-mechanical oscillators, levitated in vacuum. Each oscillator consists of a polystyrene microsphere trapped in a circularly polarized, counter-propagating Gaussian laser beam. Non-conservative, azimuthal forces, deriving from inhomogeneous optical spin, push the micro-particles out of thermodynamic equilibrium. For modest optical powers each particle shows a tendency towards orbital circulation. Initially, their stochastic motion is weakly correlated. As the power is increased, the tendency towards orbital circulation strengthens and the motion of the particles becomes highly correlated. Eventually, centripetal forces overcome optical gradient forces and the oscillators undergo a collective Hopf bifurcation. For laser powers exceeding this threshold, a pair of limit cycles appear, which synchronize due to weak optical and hydrodynamic interactions. In principle, arrays of such Non-Hermitian elements can be arranged, paving the way for opto-mechanical topological materials or, possibly, classical time crystals. In addition, the preparation of synchronized states in levitated optomechanics could lead to new and robust sensors or alternative routes to the entanglement of macroscopic objects.

Rapid Identification of Pathogens Causing Bloodstream Infections by Raman Spectroscopy and Raman Tweezers.

The search for the "Holy Grail" in clinical diagnostic microbiology-a reliable, accurate, low-cost, real-time, easy-to-use method-has brought up several methods with the potential to meet these criteria. One is Raman spectroscopy, an optical, nondestructive method based on the inelastic scattering of monochromatic light. The current study focuses on the possible use of Raman spectroscopy for identifying microbes causing severe, often life-threatening bloodstream infections. We included 305 microbial strains of 28 species acting as causative agents of bloodstream infections. Raman spectroscopy identified the strains from grown colonies, with 2.8% and 7% incorrectly identified strains using the support vector machine algorithm based on centered and uncentred principal-component analyses, respectively. We combined Raman spectroscopy with optical tweezers to speed up the process and captured and analyzed microbes directly from spiked human serum. The pilot study suggests that it is possible to capture individual microbial cells from human serum and characterize them by Raman spectroscopy with notable differences among different species. IMPORTANCE Bloodstream infections are among the most common causes of hospitalizations and are often life-threatening. To establish an effective therapy for a patient, the timely identification of the causative agent and characterization of its antimicrobial susceptibility and resistance profiles are essential. Therefore, our multidisciplinary team of microbiologists and physicists presents a method that reliably, rapidly, and inexpensively identifies pathogens causing bloodstream infections-Raman spectroscopy. We believe that it might become a valuable diagnostic tool in the future. Combined with optical trapping, it offers a new approach where the microorganisms are individually trapped in a noncontact way by optical tweezers and investigated by Raman spectroscopy directly in a liquid sample. Together with the automatic processing of measured Raman spectra and comparison with a database of microorganisms, it makes the whole identification process almost real time.

SERS-Tags: Selective Immobilization and Detection of Bacteria by Strain-Specific Antibodies and Surface-Enhanced Raman Scattering.

Efficient separation and sensitive identification of pathogenic bacterial strains is essential for a prosperous modern society, with direct applications in medical diagnostics, drug discovery, biodefense, and food safety. We developed a fast and reliable method for antibody-based selective immobilization of bacteria from suspension onto a gold-plated glass surface, followed by detection using strain-specific antibodies linked to gold nanoparticles decorated with a reporter molecule. The reporter molecules are subsequently detected by surface-enhanced Raman spectroscopy (SERS). Such a multi-functionalized nanoparticle is called a SERS-tag. The presented procedure uses widely accessible and cheap materials for manufacturing and functionalization of the nanoparticles and the immobilization surfaces. Here, we exemplify the use of the produced SERS-tags for sensitive single-cell detection of opportunistic pathogen Escherichia coli, and we demonstrate the selectivity of our method using two other bacterial strains, Staphylococcus aureus and Serratia marcescens, as negative controls. We believe that the described approach has a potential to inspire the development of novel medical diagnostic tools for rapid identification of bacterial pathogens.

Rapid identification of pathogens in blood serum via Raman tweezers in combination with advanced processing methods.

Pathogenic microbes contribute to several major global diseases that kill millions of people every year. Bloodstream infections caused by these microbes are associated with high morbidity and mortality rates, which are among the most common causes of hospitalizations. The search for the "Holy Grail" in clinical diagnostic microbiology, a reliable, accurate, low cost, real-time, and easy-to-use diagnostic method, is one of the essential issues in clinical practice. These very critical conditions can be met by Raman tweezers in combination with advanced analysis methods. Here, we present a proof-of-concept study based on Raman tweezers combined with spectral mixture analysis that allows for the identification of microbial strains directly from human blood serum without user intervention, thus eliminating the influence of a data analyst.

In Vivo Contrast Imaging of Rat Heart with Carbon Dioxide Foam.

Widely used classical angiography with the use of iodine contrast agents is highly problematic, particularly in patients with diabetes mellitus, cardiac and pulmonary diseases, or degree III or IV renal insufficiency. Some patients may be susceptible to allergic reaction to the iodine contrast substance. The intravenous injection of a bolus of CO2 (negative contrast) is an alternative method, which is, however, currently only used for imaging blood vessels of the lower limbs. The aim of our project was to design and test on an animal model a methodology for injecting the CO2 foam which would minimize the possibility of embolization of the brain tissue and heart infarction, leading to their damage. This is important research for the further promotion of the use of CO2, which is increasingly important for endovascular diagnosis and treatment, because carbon-dioxide-related complications are extremely rare. CO2 foam was prepared by the rapid mixing in a 2:1 ratio of CO2 and fetal bovine serum (FBS)-enriched Dulbecco's Modified Eagle Medium (DMEM). Freshly prepared CO2 foam was administered into the catheterized rat tail vein or cannulated rat abdominal aorta and inferior vena cava (IVC). CO2 foam was compared with commercially available microbubbles (lipid shell/gas core). The rat heart in its parasternal long axis was imaged in B-Mode and Non-linear Contrast Mode before/during and after the contrast administration. Samples of the brain, heart and lungs were collected and subjected to histological examination. The non-linear contrast imaging method enables the imaging of micron-sized gas microbubbles inside a rat heart. The significantly shorter lifetime of the prepared CO2 foam is a benefit for avoiding the local ischemia of tissues.

Raman spectroscopy-a tool for rapid differentiation among microbes causing urinary tract infections.

Urinary tract infections belong to the most common infections in the world. Besides community-acquired infections, nosocomial infections pose a high risk especially for patients having indwelling catheters, undergoing urological surgeries or staying at hospital for prolonged time. They can be often complicated by antimicrobial resistance and/or biofilm formation. Therefore, a rapid diagnostic tool enabling timely identification of a causative agent and its susceptibility to antimicrobials is a need. Raman spectroscopy appears to be a suitable method that allows rapid differentiation among microbes and provides a space for further analyses, such as determination of capability of biofilm formation or antimicrobial susceptibility/resistance in tested strains. Our work here presents a possibility to differ among most common microbes causing urinary tract infections (belonging to 20 species). We tested 254 strains directly from colonies grown on Mueller-Hinton agar plates. The results show that it is possible to distinguish among the tested species using Raman spectroscopy, which proves its great potential for future use in clinical diagnostics. Moreover, we present here a pilot study of a real-time analysis and identification (in less than 10 min) of single microbial cells directly in urine employing optical tweezers combined with Raman spectroscopy.

Challenges on optical printing of colloidal nanoparticles.

While colloidal chemistry provides ways to obtain a great variety of nanoparticles with different shapes, sizes, material compositions, and surface functions, their controlled deposition and combination on arbitrary positions of substrates remain a considerable challenge. Over the last ten years, optical printing arose as a versatile method to achieve this purpose for different kinds of nanoparticles. In this article, we review the state of the art of optical printing of single nanoparticles and discuss its strengths, limitations, and future perspectives by focusing on four main challenges: printing accuracy, resolution, selectivity, and nanoparticle photostability.

Optically Transportable Optofluidic Microlasers with Liquid Crystal Cavities Tuned by the Electric Field.

Liquid crystal microdroplets with readily adjustable optical properties have attracted considerable attention for building reconfigurable optofluidic microsystems for sensing, imaging, and light routing applications. In this quest, development of active optical microcavities serving as versatile integrated sources of coherent light and ultra-sensitive environmental sensors has played a prominent role. Here, we study transportable optofluidic microlasers reversibly tunable by an external electric field, which are based on fluorophore-doped emulsion droplets of radial nematic liquid crystals manipulated by optical tweezers in microfluidic chips with embedded liquid electrodes. Full transparency of the electrodes formed by a concentrated electrolyte solution allows for applying an electric field to the optically trapped droplets without undesired heating caused by light absorption. Taking advantage of independent, precise control over the electric and thermal stimulation of the lasing liquid crystal droplets, we characterize their spectral tuning response at various optical trapping powers and study their relaxation upon a sudden decrease in the trapping power. Finally, we demonstrate that sufficiently strong applied electric fields can induce fully reversible phase transitions in the trapped droplets even below the bulk melting temperature of the used liquid crystal. Our observations indicate viability of creating electrically tunable, optically transported microlasers that can be prepared on-demand and operated within microfluidic chips to implement integrated microphotonic or sensing systems.

Raman Microspectroscopic Analysis of Selenium Bioaccumulation by Green Alga Chlorella vulgaris.

Selenium (Se) is an element with many commercial applications as well as an essential micronutrient. Dietary Se has antioxidant properties and it is known to play a role in cancer prevention. However, the general population often suffers from Se deficiency. Green algae, such as Chlorella vulgaris, cultivated in Se-enriched environment may be used as a food supplement to provide adequate levels of Se. We used Raman microspectroscopy (RS) for fast, reliable, and non-destructive measurement of Se concentration in living algal cells. We employed inductively coupled plasma-mass spectrometry as a reference method to RS and we found a substantial correlation between the Raman signal intensity at 252 cm-1 and total Se concentration in the studied cells. We used RS to assess the uptake of Se by living and inactivated algae and demonstrated the necessity of active cellular transport for Se accumulation. Additionally, we observed the intracellular Se being transformed into an insoluble elemental form, which we further supported by the energy-dispersive X-ray spectroscopy imaging.

Complex colloidal structures with non-linear optical properties formed in an optical trap.

Illumination of a colloidal suspension of dielectric nanoparticles (50 nm in radius) with counter-propagating non-interfering laser beams of sufficient power leads to spatial redistribution of particles due to associated optical forces and formation of colloidal structures composed of thousands of nanoparticles along the beams. We employ a weak probe beam propagating through the colloidal structure and demonstrate that the colloidal structure acts effectively as a non-linear optical medium, similar to a gradient index lens, with optical transformation properties externally tunable by trapping laser power. With an increasing number of nanoparticles we observe the formation of a more complex colloidal structure axially and even laterally and we explain the origin of this process.

Using the transient trajectories of an optically levitated nanoparticle to characterize a stochastic Duffing oscillator.

We propose a novel methodology to estimate parameters characterizing a weakly nonlinear Duffing oscillator represented by an optically levitating nanoparticle. The method is based on averaging recorded trajectories with defined initial positions in the phase space of nanoparticle position and momentum and allows us to study the transient dynamics of the nonlinear system. This technique provides us with the parameters of a levitated nanoparticle such as eigenfrequency, damping, coefficient of nonlinearity and effective temperature directly from the recorded transient particle motion without any need for external driving or modification of an experimental system. Comparison of this innovative approach with a commonly used method based on fitting the power spectrum density profile shows that the proposed complementary method is applicable even at lower pressures where the nonlinearity starts to play a significant role and thus the power spectrum density method predicts steady state parameters. The technique is applicable also at low temperatures and extendable to recent quantum experiments. The proposed method is applied on experimental data and its validity for one-dimensional and three-dimensional motion of a levitated nanoparticle is verified by extensive numerical simulations.

Analysis of Bacteriophage-Host Interaction by Raman Tweezers.

Bacteriophages, or "phages" for short, are viruses that replicate in bacteria. The therapeutic and biotechnological potential of phages and their lytic enzymes is of interest for their ability to selectively destroy pathogenic bacteria, including antibiotic-resistant strains. Introduction of phage preparations into medicine, biotechnology, and food industry requires a thorough characterization of phage-host interaction on a molecular level. We employed Raman tweezers to analyze the phage-host interaction of Staphylococcus aureus strain FS159 with a virulent phage JK2 (=812K1/420) of the Myoviridae family and a temperate phage 80α of the Siphoviridae family. We analyzed the timeline of phage-induced molecular changes in infected host cells. We reliably detected the presence of replicating phages in bacterial cells within 5 min after infection. Our results lay the foundations for building a Raman-based diagnostic instrument capable of real-time, in vivo, in situ, nondestructive characterization of the phage-host relationship on the level of individual cells, which has the potential of importantly contributing to the development of phage therapy and enzybiotics.

Nanovortex-Driven All-Dielectric Optical Diffusion Boosting and Sorting Concept for Lab-on-a-Chip Platforms.

The ever-growing field of microfluidics requires precise and flexible control over fluid flows at reduced scales. Current constraints demand a variety of controllable components to carry out several operations inside microchambers and microreactors. In this context, brand-new nanophotonic approaches can significantly enhance existing capabilities providing unique functionalities via finely tuned light-matter interactions. A concept is proposed, featuring dual on-chip functionality: boosted optically driven diffusion and nanoparticle sorting. High-index dielectric nanoantennae is specially designed to ensure strongly enhanced spin-orbit angular momentum transfer from a laser beam to the scattered field. Hence, subwavelength optical nanovortices emerge driving spiral motion of plasmonic nanoparticles via the interplay between curl-spin optical forces and radiation pressure. The nanovortex size is an order of magnitude smaller than that provided by conventional beam-based approaches. The nanoparticles mediate nanoconfined fluid motion enabling moving-part-free nanomixing inside a microchamber. Moreover, exploiting the nontrivial size dependence of the curled optical forces makes it possible to achieve precise nanoscale sorting of gold nanoparticles, demanded for on-chip separation and filtering. Altogether, a versatile platform is introduced for further miniaturization of moving-part-free, optically driven microfluidic chips for fast chemical analysis, emulsion preparation, or chemical gradient generation with light-controlled navigation of nanoparticles, viruses or biomolecules.

Coherent oscillations of a levitated birefringent microsphere in vacuum driven by nonconservative rotation-translation coupling.

We demonstrate an effect whereby stochastic, thermal fluctuations combine with nonconservative optical forces to break detailed balance and produce increasingly coherent, apparently deterministic motion for a vacuum-trapped particle. The particle is birefringent and held in a linearly polarized Gaussian optical trap. It undergoes oscillations that grow rapidly in amplitude as the air pressure is reduced, seemingly in contradiction to the equipartition of energy. This behavior is reproduced in direct simulations and captured in a simplified analytical model, showing that the underlying mechanism involves nonsymmetric coupling between rotational and translational degrees of freedom. When parametrically driven, these self-sustained oscillators exhibit an ultranarrow linewidth of 2.2 µHz and an ultrahigh mechanical quality factor in excess of 2 × 108 at room temperature. Last, nonequilibrium motion is seen to be a generic feature of optical vacuum traps, arising for any system with symmetry lower than that of a perfect isotropic microsphere in a Gaussian trap.

Controlled Oil/Water Partitioning of Hydrophobic Substrates Extending the Bioanalytical Applications of Droplet-Based Microfluidics.

Functional annotation of novel proteins lags behind the number of sequences discovered by the next-generation sequencing. The throughput of conventional testing methods is far too low compared to sequencing; thus, experimental alternatives are needed. Microfluidics offer high throughput and reduced sample consumption as a tool to keep up with a sequence-based exploration of protein diversity. The most promising droplet-based systems have a significant limitation: leakage of hydrophobic compounds from water compartments to the carrier prevents their use with hydrophilic reagents. Here, we present a novel approach of substrate delivery into microfluidic droplets and apply it to high-throughput functional characterization of enzymes that convert hydrophobic substrates. Substrate delivery is based on the partitioning of hydrophobic chemicals between the oil and water phases. We applied a controlled distribution of 27 hydrophobic haloalkanes from oil to reaction water droplets to perform substrate specificity screening of eight model enzymes from the haloalkane dehalogenase family. This droplet-on-demand microfluidic system reduces the reaction volume 65 000-times and increases the analysis speed almost 100-fold compared to the classical test tube assay. Additionally, the microfluidic setup enables a convenient analysis of dependences of activity on the temperature in a range of 5 to 90 °C for a set of mesophilic and hyperstable enzyme variants. A high correlation between the microfluidic and test tube data supports the approach robustness. The precision is coupled to a considerable throughput of >20 000 reactions per day and will be especially useful for extending the scope of microfluidic applications for high-throughput analysis of reactions including compounds with limited water solubility.

Identification of ability to form biofilm in Candida parapsilosis and Staphylococcus epidermidis by Raman spectroscopy.

Aim: Finding rapid, reliable diagnostic methods is a big challenge in clinical microbiology. Raman spectroscopy is an optical method used for multiple applications in scientific fields including microbiology. This work reports its potential in identifying biofilm positive strains of Candida parapsilosis and Staphylococcus epidermidis. Materials & methods: We tested 54 S. epidermidis strains (23 biofilm positive, 31 negative) and 51 C. parapsilosis strains (27 biofilm positive, 24 negative) from colonies on Mueller-Hinton agar plates, using Raman spectroscopy. Results: The accuracy was 98.9% for C. parapsilosis and 96.1% for S. epidermidis. Conclusion: The method showed great potential for identifying biofilm positive bacterial and yeast strains. We suggest that Raman spectroscopy might become a useful aid in clinical diagnostics.

Spin to orbital light momentum conversion visualized by particle trajectory.

In a tightly focused beam of light having both spin and orbital angular momentum, the beam exhibits the spin-orbit interaction phenomenon. We demonstrate here that this interaction gives rise to series of subtle, but observable, effects on the dynamics of a dielectric microsphere trapped in such a beam. In our setup, we control the strength of spin-orbit interaction with the width, polarization and vorticity of the beam and record how these parameters influence radius and orbiting frequency of the same single orbiting particle pushed by the laser beam. Using Richard and Wolf model of the non-paraxial beam focusing, we found a very good agreement between the experimental results and the theoretical model based on calculation of the optical forces using the generalized Lorenz-Mie theory extended to a non-paraxial vortex beam. Especially the radius of the particle orbit seems to be a promising parameter characterizing the spin to orbital momentum conversion independently on the trapping beam power.

Optomechanical properties of optically self-arranged colloidal waveguides.

When a suspension of wavelength-sized polystyrene spheres is illuminated with non-interfering counter-propagating Gaussian beams, the particles self-arrange into a colloidal waveguide (CWG). Mutual force interaction among particles is mediated by scattered light, referred to as the optical binding. We analyzed the longitudinal and lateral motion of particles in such CWGs made of an increasing number of particles with diameters of either 520 or 657 nm. We observed the enhancement of the binding stiffness of neighboring particles by more than an order of magnitude. This enhancement is done by optical means, mainly due to a local increase of optical intensity due to multiple light scattering in an optically bound structure.