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Aeromonas salmonicida is the pathogen of furunculosis, causing a septicaemic infection that influences both salmonid and non-salmonid fish. Early diagnosis of these contagions is essential for disease surveillance and prevention, so that the rapid and sensitive approach is required. Herein, recombinase polymerase amplification (RPA) assay and RPA with a lateral flow dipstick (RPA-LFD) were produced for testing A. salmonicida. The RPA and RPA-LFD primer sets were devised based on the conserved fragment sequence of vapA gene. Then, RPA and RPA-LFD reaction systems were established, and the reaction temperature and time were optimized. After optimization, the RPA method was capable to test A. salmonicida within 10 min and the RPA-LFD method can detect A. salmonicida in only 5 min. The RPA and RPA-LFD methods exhibited high specificity with no cross-reaction with others strains. To assess sensitivity, a partial vapA gene was cloned, and serial plasmid dilutions were created ranging from 1×106 to 1×10− 1 copies/µL. The detection limit of RPA was 1×102 copies/µL and the detection limit of RPA-LFD was 1 copies/µL. For spiked turbot tissue samples, the sensitivity detecting of A. salmonicida was 1.2×101 CFU/mL and 1.2 CFU/mL by RPA and RPA-LFD, respectively. Therefore, our RPA and RPA-LFD assays exhibited excellent specificity and sensitivity, which provided two simple, fast and dependable methods to conduct large-scale field investigations of A. salmonicida in resource-limited settings.

In this research, Vibrio vulnificus formalin-killed (FKCs) vaccine and ghost (VVGs) vaccine were successfully developed, and shown to prevent vibriosis of Scophthalmus maximus resulting from V. vulnificus. The antibody titre of FKCs and VVGs vaccine was 1: 2

Aeromonas salmonicida is the pathogen underlying furunculosis, causing a septicemic infection that influences both salmonid and non-salmonid fish. Early diagnosis of these contagions is essential for disease surveillance and prevention, so a rapid and sensitive approach is needed. Herein, a recombinase polymerase amplification EXO (RPA-EXO) assay and RPA with a lateral flow dipstick (RPA-LFD) were produced for testing A. salmonicida. The RPA-EXO and RPA-LFD primer sets were devised based on the conserved fragment sequence of the vapA gene. Then, RPA-EXO and RPA-LFD reaction systems were established, and the reaction temperature and time were optimized. After optimization, the RPA-EXO method was capable of testing A. salmonicida within 10 min, and the RPA-LFD method could detect A. salmonicida in only 5 min. The RPA-EXO and RPA-LFD methods exhibited high specificity with no cross-reaction with other strains. To assess sensitivity, a partial vapA gene was cloned, and serial plasmid dilutions were created ranging from 1 × 10

Single-shot ultrafast multi-frame imaging technology plays a crucial role in the observation of laser-induced plasma. However, there are many challenges in the application of laser processing, such as technology fusion and imaging stability. To provide a stable and reliable observation method, we propose an ultrafast single-shot multi-frame imaging technology based on wavelength polarization multiplexing. Through the frequency doubling and birefringence effects of the BBO and the quartz crystal, the 800 nm femtosecond laser pulse was frequency doubled to 400 nm, and a sequence of probe sub-pulses with dual-wavelength and different polarization was generated. The coaxial propagation and framing imaging of multi-frequency pulses provided stable imaging quality and clarity, as well as high temporal/spatial resolution (200 fs and 228 lp/mm). In the experiments involving femtosecond laser-induced plasma propagation, the probe sub-pulses measured their time intervals by capturing the same results. Specifically, the measured time intervals were 200 fs between the same color pulses and 1 ps between the adjacent different. Finally, based on the obtained system time resolution, we observed and revealed the evolution mechanism of femtosecond laser-induced air plasma filaments, the multifilament propagation of femtosecond laser in fused silica, and the influence mechanism of air ionization on laser-induced shock waves.

The femtosecond laser ablation of metals is a complex and violent nonequilibrium process, and numerous studies have sought to reveal the evolution of a single physical phenomenon, such as laser-induced periodic surface micro-nanostructures or plasma eruptions. By considering the multiphysics scenarios of energy and heat transfer, structural mechanics, hydrodynamics, and nucleation dynamics, a femto-nanosecond and nano-micrometer multiscale framework combining electron–phonon-coupled heat transfer, lattice deformation, phase transition, and plasma eruption was constructed to describe the heat and mass transfer mechanism of femtosecond laser–copper interaction. A multiphysics model was proposed in this study to simulate the ablation process with different laser fluences. Ablation occurs at low near-threshold fluences primarily via a combination of the thermal phase transition process of melting and thermoplastic deformation coupled with the nonthermal phase transition process of hot electron explosion. Marangoni convection and non-uniform nucleation at the solid–liquid interface create micro-nano structures on the surface of the ablation crater. At a high laser fluence, plasma plumes are emitted via gasification and eruption, and as the material is heated to decrease its density, the surface is broken into a micro-column structure, and then the micro-columns fracture and erupt to form micro-nano structures and plasma plumes. Numerical results offer a better understanding of surface topography modifications and plasma plume evolution and promote the application of femtosecond laser precision fabrication in the fields of aviation, mechanics, electronics, and materials engineering.

Topological insulators are a new class of quantum materials with metallic (edge) surface states and insulating bulk states. They exhibit various novel electronic and optical properties that make them highly promising electronic, spintronic, and optoelectronic materials. Our report confirms that the topological insulator Bi 1.5 Sb 0.5 Te 1.8 Se 1.2 (BSTS) is also an effective plasmonic material in the visible and near-infrared range. A BSTS film can effectively control transmission and reflection characteristics by changing the period of the hole array. This study determined that a strong resonant surface plasmonic mode at the resonance peak can confine approximately 80% of the electromagnetic field energy is demonstrated. Higher-order (second- and third-order) resonance peaks were also found, which is critical for controlling electromagnetic waves and research into new optoelectronic devices.

In this study, femtosecond laser double pulses were tested to improve their nickel ablation efficiency. The experimental results indicated that compared with single pulses, double pulses with different delay times generated craters with larger diameters and depths. The results obtained for three sets of double pulses with different energy ratios indicated that double pulses with an energy ratio of 1:9 had the highest ablation efficiency, followed by those with energy ratios of 2:8 and 5:5. The double pulses with the aforementioned three energy ratios achieved the maximum ablation efficiency when the delay time was 3–4 ps. Compared with single pulses, double pulses with an energy ratio of 1:9 generated craters with an up to 34% greater depth and up to 14% larger diameter. In addition, an interference effect was observed with a double pulse delay time of 0 ps, which has seldom been reported in the literature. The double pulses were simulated using the two-temperature model. The simulation results indicated that double pulses with an energy ratio of 1:9 with a delay time of 4 ps can perform the strongest ablation. These simulation results are in line with the experimental results.

Femtosecond laser fabrication has grown to be a major method of extreme manufacturing because of the extreme energy density and spatial and temporal scales of femtosecond lasers. The physical effects and the mechanism of interaction between femtosecond lasers and materials are distinct from those in traditional processes. The nonlinear and nonequilibrium effects of the interaction have given rise to new concepts, principles, and methods, such as femtosecond pulse durations are shorter than many physical/chemical characteristic times, which permits manipulating, adjusting, or interfering with electron dynamics. These new concepts and methods have broad application prospects in micro/nanofabrication, chemical synthesis, material processing, quantum control, and other related fields. This review discusses the cutting-edge theories, methods, measurements, and applications of femtosecond lasers to micro/nano-manufacturing. The key to future development of femtosecond laser manufacturing lies in revealing its fabrication mechanism from the electronic level and precisely regulating the electronic dynamics.

Imaging flow cytometry is a powerful tool by virtue of its capability for high-throughput cell analysis. The advent of high-speed optical imaging methods on a microfluidic platform has significantly improved cell throughput and brought many degrees of freedom to instrumentation and applications over the last decade, but it also poses a predicament on microfluidic chips. Specifically, as the throughput increases, the flow speed also increases (currently reaching 10 m/s): consequently, the increased hydrodynamic pressure on the microfluidic chip deforms the wall of the microchannel and produces detrimental effects lead to defocused and blur image. Here, we present a comprehensive study of the effects of flow-induced microfluidic chip wall deformation on imaging flow cytometry. We fabricated three types of microfluidic chips with the same geometry and different degrees of stiffness made of polydimethylsiloxane (PDMS) and glass to investigate material influence on image quality. First, we found the maximum deformation of a PDMS microchannel was >60 μm at a pressure of 0.6 MPa, while no appreciable deformation was identified in a glass microchannel at the same pressure. Second, we found the deviation of lag time that indicating velocity difference of migrating microbeads due to the deformation of the microchannel was 29.3 ms in a PDMS microchannel and 14.9 ms in a glass microchannel. Third, the glass microchannel focused cells into a slightly narrower stream in the X-Y plane and a significantly narrower stream in the Z-axis direction (focusing percentages were increased 30%, 32%, and 5.7% in the glass channel at flow velocities of 0.5, 1.5, and 3 m/s, respectively), and the glass microchannel showed stabler equilibrium positions of focused cells regardless of flow velocity. Finally, we achieved the world's fastest imaging flow cytometry by combining a glass microfluidic device with an optofluidic time-stretch microscopy imaging technique at a flow velocity of 25 m/s. © 2019 International Society for Advancement of Cytometry.

Dielectric metasurfaces can achieve flexible beam manipulations. Herein, we study dielectric metasurfaces with different refractive indices, periods, incident angles, and cross-sectional shapes to determine the metasurface working mechanisms. Perfect transmission mainly depends on multipolar interference that can be used to control the transmission modes through the hybrid periods, hybrid cross sections, and multilayers. Perfect reflection is strongly influenced by the period of the metasurface and occurs only when the period is shorter than incident wavelength, which can be attributed to the lattice coupling. Furthermore, lattice coupling can be classified into two types with distinct properties: vertical mode and horizontal mode coupling. The vertical mode appears when the effective wavelength matches the feature size, whereas the horizontal mode only appears when the incident wavelength is close to the period. The horizontal mode is sensitive to the incident angle. The revealed functioning mechanisms enable further practical applications of metasurfaces.

Recently, the application of femtosecond laser ablation of nickel to prepare super-hydrophobic structures in marine anticorrosion has become more and more mature, but the processing efficiency is slow, which limits its large-scale application. In this study, a method for improving the efficiency of ablating nickel through multiple scanning of femtosecond lasers single pulse is proposed. The experimental results show that when the total processing time is the same, the higher scanning speed and more scanning times will promote the increase of the groove depth formed by the ablated nickel compared with the low scanning speed. This phenomenon has been verified under three different laser fluences. Compared with 25 um/s, the ablation efficiency is increased by 42.6%, 48.2% and 55.4% when scanning speed is 200 um/s, respectively. The difference in ablation efficiency in the experiment is related to the fact that femtosecond laser ablation of nickel will change the physical and chemical properties of the bottom of the material.

Cell sorting requires a stable flowing speed, especially for a fast flow cytometry like the optical time-stretch imaging. The flowing speed fluctuation of Euglena cells and Nile red particles is tested and evaluated by a home-built flowing speed testing system. Higher flowing speeds have better stability. Flowing microchannels with controlling steps also improve the speed stability. The time fluctuation is ~ 1 ms under a flowing speed of ~ 1.4 m/s. Cell sorting can be realized by either using an ultra-short delay time between the sorting point and imaging point, which is suitable for relative lower cells counting speed (~ 5000 cells/s), or developing effective fluctuation controlling methods for higher counting speed, like using step glass microchannels.

Defect/active site control is crucial for tuning the chemical, optical, and electronic properties of MoS2, which can adjust the performance of MoS2 in application areas such as electronics, optics, catalysis, and molecular sensing. This study presents an effective method of inducing defect/active sites, including micro/nanofractured structures and S atomic vacancies, on monolayer MoS2 flakes by using femtosecond laser pulses, through which physical–chemical adsorption and charge transfer between foreign molecules (O2 or R6G molecules) and MoS2 are enhanced. The enhanced charge transfer between foreign molecules (O2 or R6G) and femtosecond laser-treated MoS2 can enhance the electronic doping effect between them, hence resulting in a photoluminescence photon energy shift (reaching 0.05 eV) of MoS2 and Raman enhancement (reaching 6.4 times) on MoS2 flakes for R6G molecule detection. Finally, photoluminescence control and micropatterns on MoS2 and surface-enhanced-Raman-scattering (SERS) enhancement of MoS2 for organic molecule detection are achieved. The proposed method, which can control the photoluminescence properties and arbitrary micropatterns on MoS2 and enhance its chemicobiological sensing performance for organic/biological molecules, has advantages of simplicity, maskless processing, strong controllability, high precision, and high flexibility, highlighting the superior ability of femtosecond laser pulses to achieve the property control and functionalization of two-dimensional materials.

The evolution of CaO-Al2O3-MgO-(SiO2) inclusions associated with CaS during electroslag rapid remelting (ESRR) of the tool steel with ultra low oxygen (8 ppm) under protective atmosphere was investigated. Thermodynamic calculation was performed to evaluate the oxygen level of liquid steel and the inclusion evolution during ESRR with the help of FactSage software (CON2 database). The results demonstrated that the reoxidation of liquid steel took place during industrial ESRR, resulting in the oxygen pickup of approximately double content. The oxygen level in the steel was determined by [Al]-[O] equilibrium during ESRR, whereas FeO in the slag transferred oxygen into the liquid steel, as indicated by the significant oxygen pickup. The inclusions in the steel electrode are oxide–sulfide type of CaO-Al2O3-MgO partially wrapped with CaS. The original oxide inclusions that had not been removed in ESRR process were transformed to liquid CaO-Al2O3-SiO2-MgO inclusions through reacting with the dissolved oxygen supplied from the reoxidation of liquid steel and concerned elements in liquid steel during ESRR. These CaO-Al2O3-SiO2-MgO inclusions were invariably associated with patch-type or shell-type CaS, which hold different formation mechanisms. The inclusions ranging from 2 to 4 µm take up a dominant proportion (a little more than 50 pct of the total inclusions) in the remelted ingots, and those larger than 8 µm account for about 3 pct. Melting rates of ESRR not only exerted a negligible effect on the steel cleanliness and the removal efficiency of oxide inclusions, but also on the inclusion size distribution.

Innovations in optical microscopy have opened new windows onto scientific research, industrial quality control, and medical practice over the last few decades. One of such innovations is optofluidic time-stretch quantitative phase microscopy - an emerging method for high-throughput quantitative phase imaging that builds on the interference between temporally stretched signal and reference pulses by using dispersive properties of light in both spatial and temporal domains in an interferometric configuration on a microfluidic platform. It achieves the continuous acquisition of both intensity and phase images with a high throughput of more than 10,000 particles or cells per second by overcoming speed limitations that exist in conventional quantitative phase imaging methods. Applications enabled by such capabilities are versatile and include characterization of cancer cells and microalgal cultures. In this paper, we review the principles and applications of optofluidic time-stretch quantitative phase microscopy and discuss its future perspective.

The development of reliable, sustainable, and economical sources of alternative fuels to petroleum is required to tackle the global energy crisis. One such alternative is microalgal biofuel, which is expected to play a key role in reducing the detrimental effects of global warming as microalgae absorb atmospheric CO2 via photosynthesis. Unfortunately, conventional analytical methods only provide population-averaged lipid amounts and fail to characterize a diverse population of microalgal cells with single-cell resolution in a non-invasive and interference-free manner. Here high-throughput label-free single-cell screening of lipid-producing microalgal cells with optofluidic time-stretch quantitative phase microscopy was demonstrated. In particular, Euglena gracilis, an attractive microalgal species that produces wax esters (suitable for biodiesel and aviation fuel after refinement), within lipid droplets was investigated. The optofluidic time-stretch quantitative phase microscope is based on an integration of a hydrodynamic-focusing microfluidic chip, an optical time-stretch quantitative phase microscope, and a digital image processor equipped with machine learning. As a result, it provides both the opacity and phase maps of every single cell at a high throughput of 10,000 cells/s, enabling accurate cell classification without the need for fluorescent staining. Specifically, the dataset was used to characterize heterogeneous populations of E. gracilis cells under two different culture conditions (nitrogen-sufficient and nitrogen-deficient) and achieve the cell classification with an error rate of only 2.15%. The method holds promise as an effective analytical tool for microalgae-based biofuel production. © 2017 International Society for Advancement of Cytometry.

Optical time-stretch microscopy has recently attracted intensive attention for its capability of acquiring images at an ultrahigh frame rate. Unfortunately, its achievable frame rate is limited by the requirement of having no overlap between consecutive frames, which leads to a tradeoff between the frame rate (pulse repetition rate) and the amount of the temporal dispersion used for optical image serialization. In this paper, we demonstrate compressive sensing on the platform of optical time-stretch microscopy to overcome the tradeoff between frame rate and temporal dispersion (time stretch) and achieve 50 times higher frame rate than conventional optical time-stretch microscopy. Specifically, we computationally perform compressed optical time-stretch microscopy with an experimental dataset acquired by conventional optical time-stretch microscopy and demonstrate its effects in terms of spatial resolution and cell classification accuracy. Our results indicate that the spatial resolution and cell classification accuracy reach 780 nm and 95% at a line scan rate of 675 MHz and 6.75 GHz, respectively, which correspond to five times and 50 times higher frame rates than what conventional optical time-stretch microscopy can achieve with the same dispersion amount and digitizer sampling rate.

Drug susceptibility (also called chemosensitivity) is an important criterion for developing a therapeutic strategy for various cancer types such as breast cancer and leukemia. Recently, functional assays such as high-content screening together with genomic analysis have been shown to be effective for predicting drug susceptibility, but their clinical applicability is poor since they are time-consuming (several days long), labor-intensive, and costly. Here we present a highly simple, rapid, and cost-effective liquid biopsy for ex vivo drug-susceptibility testing of leukemia. The method is based on an extreme-throughput (>1 million cells per second), label-free, whole-blood imaging flow cytometer with a deep convolutional autoencoder, enabling image-based identification of the drug susceptibility of every single white blood cell in whole blood within 24 hours by simply flowing a drug-treated whole blood sample as little as 500 μL into the imaging flow cytometer without labeling. Our results show that the method accurately evaluates the drug susceptibility of white blood cells from untreated patients with acute lymphoblastic leukemia. Our method holds promise for affordable precision medicine.

Deep subwavelength manipulation of terahertz (THz) wave is a key method to realize compact on chip THz devices. It is demonstrated that the refractive index change in a deep subwavelength region of a dielectric layer can effectively manipulate the surface THz wave propagation by the simulation study. The feature size of this area is only 10 μm (∼1/8 λ). A slight change of refractive index, position or size of this region is enough to manipulate the surface THz waves with high efficiency, such as the transmissivity or reflectivity of different THz frequencies. Moreover, the change of the deep subwavelength region can be controlled by an ultrafast laser to achieve ultrafast dynamic manipulation of THz waves. This is a concise and efficient method of manipulating electromagnetic waves on the deep subwavelength scale and to fabricate more compact integrated optical devices.

Defect/active site control is crucial for tuning the chemical, optical, and electronic properties of MoS2, which can adjust the performance of MoS2 in application areas such as electronics, optics, catalysis, and molecular sensing. This study presents an effective method of inducing defect/active sites, including micro/nanofractured structures and S atomic vacancies, on monolayer MoS2 flakes by using femtosecond laser pulses, through which physical-chemical adsorption and charge transfer between foreign molecules (O2 or R6G molecules) and MoS2 are enhanced. The enhanced charge transfer between foreign molecules (O2 or R6G) and femtosecond laser-treated MoS2 can enhance the electronic doping effect between them, hence resulting in a photoluminescence photon energy shift (reaching 0.05 eV) of MoS2 and Raman enhancement (reaching 6.4 times) on MoS2 flakes for R6G molecule detection. Finally, photoluminescence control and micropatterns on MoS2 and surface-enhanced-Raman-scattering (SERS) enhancement of MoS2 for organic molecule detection are achieved. The proposed method, which can control the photoluminescence properties and arbitrary micropatterns on MoS2 and enhance its chemicobiological sensing performance for organic/biological molecules, has advantages of simplicity, maskless processing, strong controllability, high precision, and high flexibility, highlighting the superior ability of femtosecond laser pulses to achieve the property control and functionalization of two-dimensional materials.

Euglena gracilis (E. gracilis) has recently been attracting attention as a potential renewable source for the production of biofuels, livestock feed, cosmetics, and dietary supplements. Research has focused on strain isolation, productivity improvement, nutrient and resource allocation, and co-product production, key steps that ultimately determine the economic viability and compatibility of the biomass produced. To achieve these characteristics, approaches to select E. gracilis mutants with desirable properties, such as high wax ester content, high growth rate, and high environmental tolerance for biodiesel and biomass production, are needed. Flow-based analysis and sorting can be rapid and highly automated but calls for techniques that can precisely control the position of E. gracilis with varying sizes and shapes in a tightly focused stream in a high-throughput manner. In this work, we use a stepped microchannel consisting of a low-aspect-ratio straight channel and a series of expansion regions along the channel height. We study horizontal and vertical focusing, orientation, rotational, and translational behaviors of E. gracilis as a function of aspect ratio (AR) and channel Reynolds number (Re). By making use of inertial focusing and local secondary flows, E. gracilis with diverse shapes are directed to a single equilibrium position in a single focal stream. As an application of on-chip flow cytometry, we integrate a focusing microchip with a custom laser-two-focus (L2F) optical system and demonstrate the detection of chlorophyll autofluorescence as well as the measurement of the velocity of E. gracilis cells flowing through the microchannel.

Graded and partial graded period grating structures are proposed to trap and slow terahertz (THz) waves. Different frequencies of THz waves can be trapped at different positions along the graded grating. The exact trapping positions of every different frequency can be estimated from a fitting curve. The separation distance between trapping point of every 0.1 THz is about 1 ~ 2 mm, which can be applied in a THz waves spectrometer. An ultraslow THz waveguide can be realized by using a partial graded grating. The propagated speed of THz waves can be designed and tuned by adjusting the period of the grating. This structure offers the advantage of reducing the speed of the THz waves and propagation them over an ultra-wide spectral band. The physical trapping mechanism of the THz waves is attributed to the transformation from surface modes to cavity modes. The localized cavity modes at the trapping position have a saturated stable state. The extra energy will be reflected back along the graded grating.

In the last decade, high-content screening based on multivariate single-cell imaging has been proven effective in drug discovery to evaluate drug-induced phenotypic variations. Unfortunately, this method inherently requires fluorescent labeling which has several drawbacks. Here we present a label-free method for evaluating cellular drug responses only by high-throughput bright-field imaging with the aid of machine learning algorithms. Specifically, we performed high-throughput bright-field imaging of numerous drug-treated and -untreated cells (N = ~240,000) by optofluidic time-stretch microscopy with high throughput up to 10,000 cells/s and applied machine learning to the cell images to identify their morphological variations which are too subtle for human eyes to detect. Consequently, we achieved a high accuracy of 92% in distinguishing drug-treated and -untreated cells without the need for labeling. Furthermore, we also demonstrated that dose-dependent, drug-induced morphological change from different experiments can be inferred from the classification accuracy of a single classification model. Our work lays the groundwork for label-free drug screening in pharmaceutical science and industry.

We show that a high-energy electron bunch can be used to capture the instantaneous longitudinal and transverse field structures of the highly transient, microscopic, laser-excited relativistic wake with femtosecond resolution. The spatiotemporal evolution of wakefields in a plasma density up ramp is measured and the reversal of the plasma wake, where the wake wavelength at a particular point in space increases until the wake disappears completely only to reappear at a later time but propagating in the opposite direction, is observed for the first time by using this new technique.

According to WHO, about 10 million new cases of thrombotic disorders are diagnosed worldwide every year. Thrombotic disorders, including atherothrombosis (the leading cause of death in the US and Europe), are induced by occlusion of blood vessels, due to the formation of blood clots in which aggregated platelets play an important role. The presence of aggregated platelets in blood may be related to atherothrombosis (especially acute myocardial infarction) and is, hence, useful as a potential biomarker for the disease. However, conventional high-throughput blood analysers fail to accurately identify aggregated platelets in blood. Here we present an in vitro on-chip assay for label-free, single-cell image-based detection of aggregated platelets in human blood. This assay builds on a combination of optofluidic time-stretch microscopy on a microfluidic chip operating at a high throughput of 10 000 blood cells per second with machine learning, enabling morphology-based identification and enumeration of aggregated platelets in a short period of time. By performing cell classification with machine learning, we differentiate aggregated platelets from single platelets and white blood cells with a high specificity and sensitivity of 96.6% for both. Our results indicate that the assay is potentially promising as predictive diagnosis and therapeutic monitoring of thrombotic disorders in clinical settings.

The ability to sift through a large heterogeneous population of cells is of paramount importance in a diverse range of biomedical and green applications. Furthermore, the capability of identifying various features of cells in a label-free manner is useful for high-throughput screening. Here we present optofluidic time-stretch quantitative phase microscopy for high-throughput label-free single-cell screening. This method is based on an integration of a hydrodynamic-focusing microfluidic chip, an optical time-stretch microscope for high-speed imaging with a spatial resolution of ~800 nm at a frame rate of ~10 million frames per second, and a digital image processor for image-based characterization, classification, and statistical analysis of biological cells such as blood cells and microalgae. It provides both the opacity (amplitude) and thickness (phase) content of every cell at a high throughput of ~10,000 cells per second. This method is expected to be effective for a diverse range of applications such as cancer detection and biofuel production.

According to WHO, approximately 10 million new cases of thrombotic disorders are diagnosed worldwide every year. In the U.S. and Europe, their related diseases kill more people than those from AIDS, prostate cancer, breast cancer and motor vehicle accidents combined. Although thrombotic disorders, especially arterial ones, mainly result from enhanced platelet aggregability in the vascular system, visual detection of platelet aggregates in vivo is not employed in clinical settings. Here we present a high-throughput label-free platelet aggregate detection method, aiming at the diagnosis and monitoring of thrombotic disorders in clinical settings. With optofluidic time-stretch microscopy with a spatial resolution of 780 nm and an ultrahigh linear scanning rate of 75 MHz, it is capable of detecting aggregated platelets in lysed blood which flows through a hydrodynamic-focusing microfluidic device at a high throughput of 10,000 particles/s. With digital image processing and statistical analysis, we are able to distinguish them from single platelets and other blood cells via morphological features. The detection results are compared with results of fluorescence-based detection (which is slow and inaccurate, but established). Our results indicate that the method holds promise for real-time, low-cost, label-free, and minimally invasive detection of platelet aggregates, which is potentially applicable to detection of platelet aggregates in vivo and to the diagnosis and monitoring of thrombotic disorders in clinical settings. This technique, if introduced clinically, may provide important clinical information in addition to that obtained by conventional techniques for thrombotic disorder diagnosis, including ex vivo platelet aggregation tests.

Acquired drug resistance is a fundamental predicament in cancer therapy. Early detection of drug-resistant cancer cells during or after treatment is expected to benefit patients from unnecessary drug administration and thus play a significant role in the development of a therapeutic strategy. However, the development of an effective method of detecting drug-resistant cancer cells is still in its infancy due to their complex mechanism in drug resistance. To address this problem, we propose and experimentally demonstrate label-free image-based drug resistance detection with optofluidic time-stretch microscopy using leukemia cells (K562 and K562/ADM). By adding adriamycin (ADM) to both K562 and K562/ADM (ADM-resistant K562 cells) cells, both types of cells express unique morphological changes, which are subsequently captured by an optofluidic time-stretch microscope. These unique morphological changes are extracted as image features and are subjected to supervised machine learning for cell classification. We hereby have successfully differentiated K562 and K562/ADM solely with label-free images, which suggests that our technique is capable of detecting drug-resistant cancer cells. Our optofluidic time-stretch microscope consists of a time-stretch microscope with a high spatial resolution of 780 nm at a 1D frame rate of 75 MHz and a microfluidic device that focuses and orders cells. We compare various machine learning algorithms as well as various concentrations of ADM for cell classification. Owing to its unprecedented versatility of using label-free image and its independency from specific molecules, our technique holds great promise for detecting drug resistance of cancer cells for which its underlying mechanism is still unknown or chemical probes are still unavailable.

A graded metallic grating structure acts as a wave trapping system. Different frequencies of THz waves are trapped at different positions along this structured metal surface grating. The real wave propagation speed of such a system is reduced gradually from the light speed in vacuum to zero, which is demonstrated by calculation and simulation. Different frequencies of THz waves are propagated at a designed propagation speed by a partial graded grating according to the practical demand.

A graded metallic grating structure can not only work as a wave trapping system, but also work as an ultraslow terahertz (THz) waveguide. The depth of the grating waveguide is partial graded and partial fixed. The real propagation speed of such a system is calculated. Different frequencies of THz waves can be propagated at a designed propagation speed, even close to zero.

The development of reliable, sustainable, and economical sources of alternative fuels is an important, but challenging goal for the world. As an alternative to liquid fossil fuels, microalgal biofuel is expected to play a key role in reducing the detrimental effects of global warming since microalgae absorb atmospheric CO2 via photosynthesis. Unfortunately, conventional analytical methods only provide population-averaged lipid contents and fail to characterize a diverse population of microalgal cells with single-cell resolution in a noninvasive and interference-free manner. Here we demonstrate high-throughput label-free single-cell screening of lipid-producing microalgal cells with optofluidic time-stretch quantitative phase microscopy. In particular, we use Euglena gracilis – an attractive microalgal species that produces wax esters (suitable for biodiesel and aviation fuel after refinement) within lipid droplets. Our optofluidic time-stretch quantitative phase microscope is based on an integration of a hydrodynamic-focusing microfluidic chip, an optical time-stretch phase-contrast microscope, and a digital image processor equipped with machine learning. As a result, it provides both the opacity and phase contents of every single cell at a high throughput of 10,000 cells/s. We characterize heterogeneous populations of E. gracilis cells under two different culture conditions to evaluate their lipid production efficiency. Our method holds promise as an effective analytical tool for microalgaebased biofuel production.

The development of reliable, sustainable, and economical sources of alternative fuels to petroleum is required to tackle the global energy crisis. One such alternative is microalgal biofuel, which is expected to play a key role in reducing the detrimental effects of global warming as microalgae absorb atmospheric CO

The world is faced with environmental problems and the energy crisis due to the combustion and depletion of fossil fuels. The development of reliable, sustainable, and economical sources of alternative fuels is an important, but challenging goal for the world. As an alternative to liquid fossil fuels, algal biofuel is expected to play a key role in alleviating global warming since algae absorb atmospheric CO2 via photosynthesis. Among various algae for fuel production, Euglena gracilis is an attractive microalgal species as it is known to produce wax ester (good for biodiesel and aviation fuel) within lipid droplets. To date, while there exist many techniques for inducing microalgal cells to produce and accumulate lipid with high efficiency, few analytical methods are available for characterizing a population of such lipid-accumulated microalgae including E. gracilis with high throughout, high accuracy, and single-cell resolution simultaneously. Here we demonstrate a high-throughput optofluidic Euglena gracilis profiler which consists of an optical time-stretch microscope and a fluorescence analyzer on top of an inertial-focusing microfluidic device that can detect fluorescence from lipid droplets in their cell body and provide images of E. gracilis cells simultaneously at a high throughput of 10,000 cells/s. With the multi-dimensional information acquired by the system, we classify nitrogen-sufficient (ordinary) and nitrogen-deficient (lipid-accumulated) E. gracilis cells with a low false positive rate of 1.0%. This method provides a promise for evaluating the efficiency of lipid-inducing techniques for biofuel production, which is also applicable for identifying biomedical samples such as blood cells and cancer cells.