On-chip flow cytometer using integrated photonics for the detection of human leukocytes.

Differentiation between leukocyte subtypes like monocytes and lymphocytes is essential for cell therapy and research applications. To guarantee the cost-effective delivery of functional cells in cell therapies, billions of cells must be processed in a limited time. Yet, the sorting rates of commercial cell sorters are not high enough to reach the required yield. Process parallelization by using multiple instruments increases variability and production cost. A compact solution with higher throughput can be provided by multichannel flow cytometers combining fluidics and optics on-chip. In this work, we present a micro-flow cytometer with monolithically integrated photonics and fluidics and demonstrate that both the illumination of cells, as well as the collection of scattered light, can be realized using photonic integrated circuits. Our device is the first with sufficient resolution for the discrimination of lymphocytes and monocytes. Innovations in microfabrication have enabled complete integration of miniaturized photonic components and fluidics in a CMOS-compatible wafer stack. In combination with external optics, the device is ready for the collection of fluorescence using the on-chip excitation.

Integrated lithium niobate microwave photonic processing engine.

Integrated microwave photonics (MWP) is an intriguing technology for the generation, transmission and manipulation of microwave signals in chip-scale optical systems1,2. In particular, ultrafast processing of analogue signals in the optical domain with high fidelity and low latency could enable a variety of applications such as MWP filters3-5, microwave signal processing6-9 and image recognition10,11. An ideal integrated MWP processing platform should have both an efficient and high-speed electro-optic modulation block to faithfully perform microwave-optic conversion at low power and also a low-loss functional photonic network to implement various signal-processing tasks. Moreover, large-scale, low-cost manufacturability is required to monolithically integrate the two building blocks on the same chip. Here we demonstrate such an integrated MWP processing engine based on a 4 inch wafer-scale thin-film lithium niobate platform. It can perform multipurpose tasks with processing bandwidths of up to 67 GHz at complementary metal-oxide-semiconductor (CMOS)-compatible voltages. We achieve ultrafast analogue computation, namely temporal integration and differentiation, at sampling rates of up to 256 giga samples per second, and deploy these functions to showcase three proof-of-concept applications: solving ordinary differential equations, generating ultra-wideband signals and detecting edges in images. We further leverage the image edge detector to realize a photonic-assisted image segmentation model that can effectively outline the boundaries of melanoma lesion in medical diagnostic images. Our ultrafast lithium niobate MWP engine could provide compact, low-latency and cost-effective solutions for future wireless communications, high-resolution radar and photonic artificial intelligence.

A photonic crystal fiber-based fluorescence sensor for simultaneous and sensitive detection of lactic acid enantiomers.

A photonic crystal fiber (PCF)-based fluorescence sensor is developed for rapid and sensitive detection of lactic acid (LA) enantiomers in serum samples. The sensor is fabricated by chemical binding dual enzymes on the inner surface of the PCF with numerous pore structures and a large specific surface area, which is suitable to be utilized as an enzymatic reaction carrier. To achieve simultaneous detection of L-LA and D-LA, the PCF with an aldehyde-activated surface is cut into two separate pieces, one of which is coated with L-LDH/GPT enzymes and the other with D-LDH/GPT enzymes. By being connected and carefully aligned to each other by a suitable sleeve tube connector, the responses of both L-LA and D-LA sensors are determined by laser-induced flourescence (LIF) detection. With the aid of enzyme-linked catalytic reactions, the proposed PCF sensor can greatly improve the sensitivity and analysis speed for the detection of LA enantiomers. The PCF sensor exhibits a low limit of detection of 9.5 µM and 0.8 µM, and a wide linear range of 25-2000 µM and 2-400 µM for L-LA and D-LA, respectively. The sensor has been successfully applied to accurate determination of LA enantiomers in human serum with satisfactory reproducibility and stability. It is indicated that the present PCF sensors would be used as an attractive analytical platform for quantitative detection of trace-amount LA enantiomers in real biological samples, and thus would play a role in disease diagnosis and clinical monitoring in point-of-care testing.

Cellular Features Revealed by Transverse Laser Modes in Frequency Domain.

Biological lasers which utilize Fabry-Pérot (FP) cavities have attracted tremendous interest due to their potential in amplifying subtle biological changes. Transverse laser modes generated from cells serve as distinct fingerprints of individual cells; however, most lasing signals lack the ability to provide key information about the cell due to high complexity of transverse modes. The missing key, therefore, hinders it from practical applications in biomedicine. This study reveals the key mechanism governing the frequency distributions of transverse modes in cellular lasers. Spatial information of cells including curvature can be interpreted through spectral information of transverse modes by means of hyperspectral imaging. Theoretical studies are conducted to explore the correlation between the cross-sectional morphology of a cell and lasing frequencies of transverse modes. Experimentally, the spectral characteristics of transverse modes are investigated in live and fixed cells with different morphological features. By extracting laser modes in frequency domain, the proposed concept is applied for studying cell adhesion process and cell classification from rat cortices. This study expands a new analytical dimension of cell lasers, opening an avenue for subcellular analysis in biophotonic applications.

Label-Free Digital Detection of Intact Virions by Enhanced Scattering Microscopy.

Several applications in health diagnostics, food, safety, and environmental monitoring require rapid, simple, selective, and quantitatively accurate viral load monitoring. Here, we introduce the first label-free biosensing method that rapidly detects and quantifies intact virus in human saliva with single-virion resolution. Using pseudotype SARS-CoV-2 as a representative target, we immobilize aptamers with the ability to differentiate active from inactive virions on a photonic crystal, where the virions are captured through affinity with the spike protein displayed on the outer surface. Once captured, the intrinsic scattering of the virions is amplified and detected through interferometric imaging. Our approach analyzes the motion trajectory of each captured virion, enabling highly selective recognition against nontarget virions, while providing a limit of detection of 1 × 103 copies/mL at room temperature. The approach offers an alternative to enzymatic amplification assays for point-of-collection diagnostics.

Low-cost, thermoplastic micro-lens array with a carbon black light screen for bio-mimetic vision.

Micro-lens array is a great example of bio-mimetic technology which was inspired by compound eyes found in insects and is used in lasers, optical communication, and 3D imaging. In this study, a micro-lens array was fabricated from cyclic olefin copolymer using a cost-effective method: compression molding and thermal reflow. Also, a light screen was installed between lenses to reduce the optical interference for clearer individual images. Cyclic olefin copolymer-based micro-lens array showed good optical results under a standard optical microscope. By placing the fabricated micro-lens array directly on an image sensor, it was observed that the light screen shows significant improvement in image quality. Also, the point spread function was analyzed to confirm the optical performance and the effectiveness of the micro-lens array with the light screen installed.

Optical microlever assisted DNA stretching.

Optical microrobotics is an emerging field that has the potential to improve upon current optical tweezer studies through avenues such as limiting the exposure of biological molecules of interest to laser radiation and overcoming the current limitations of low forces and unwanted interactions between nearby optical traps. However, optical microrobotics has been historically limited to rigid, single-body end-effectors rather than even simple machines, limiting the tasks that can be performed. Additionally, while multi-body machines such as microlevers exist in the literature, they have not yet been successfully demonstrated as tools for biological studies, such as molecule stretching. In this work we have taken a step towards moving the field forward by developing two types of microlever, produced using two-photon absorption polymerisation, to perform the first lever-assisted stretches of double-stranded DNA. The aim of the work is to provide a proof of concept for using optical micromachines for single molecule studies. Both styles of microlevers were successfully used to stretch single duplexes of DNA, and the results were analysed with the worm-like chain model to show that they were in good agreement.

Light Polarization by Biological Nanocoatings.

Light plays paramount functions for living beings in nature. In addition to color, the polarization of light is used by many animals for navigation and communication. In this study, we describe the light polarizing role of special nanostructures coating cuticular surfaces of diverse arthropods. These structures are built as parallel nanoscale ridges covering the eyes of the sunlight-navigating spider Drassodes lapidosus and of the water pond-swarming black fly Simulium vittatum, as well as the light-emitting abdominal lantern of the firefly Aquatica lateralis. Exact topography and dimensions of the parallel nanoridges provide different light polarizing efficiencies and wavelength sensitivity. Optical modeling confirms that the nanoscale ridges are responsible for the spectral polarization dependency. Co-opting from our recent work on the self-assembly of Drosophila corneal nanostructures, we engineer arthropod-like parallel nanoridges on artificial surfaces, which recapitulate the light polarization effects. Our work highlights the fundamental importance of nanocoatings in arthropods for the light polarization management and provides a new biomimetic approach to produce ordered nanostructures under mild conditions.

Optically transparent microwave absorber based on water-based moth-eye structures.

We propose an approach to realize an optically transparent microwave absorber based on water-based moth-eye metamaterial structures. The absorber is made of a periodic array of properly shaped glass caps infiltrated with distilled water. Analytical calculations and numerical simulations show that the water-based metamaterial absorbs electromagnetic waves over a wide spectral band ranging from 4GHz to well above 120GHz, showing absorption levels close to 100% for incident radiation that ranges from normal to grazing angles, for both TE and TM polarizations. Yet, the structure is optically transparent, offering exciting opportunities in a variety of civil and military applications, such as for camouflage and shielding systems and in energy harvesting structures.

Microscope integrated optical coherence tomography system combined with augmented reality.

One of the disadvantages in microscope-integrated optical coherence tomography (MI-OCT) systems is that medical images acquired via different modalities are usually displayed independently. Hence, surgeons have to match two-dimensional and three-dimensional images of the same operative region subjectively. In this paper, we propose a simple registration method to overcome this problem by using guided laser points. This method combines augmented reality with an existing MI-OCT system. The basis of our idea is to introduce a guiding laser into the system, which allows us to identify fiducials in microscopic images. At first, the applied voltages of the scanning galvanometer mirror are used to calculate the fiducials' coordinates in an OCT model. After gathering data at the corresponding points' coordinates, the homography matrix and camera parameters are used to superimpose a reconstructed model on microscopic images. After performing experiments with artificial and animal eyes, we successfully obtain two-dimensional microscopic images of scanning regions with depth information. Moreover, the registration error is 0.04 mm, which is within the limits of medical and surgical errors. Our proposed method could have many potential applications in ophthalmic procedures.

Engineering Hydrogel-Based Biomedical Photonics: Design, Fabrication, and Applications.

Light guiding and manipulation in photonics have become ubiquitous in events ranging from everyday communications to complex robotics and nanomedicine. The speed and sensitivity of light-matter interactions offer unprecedented advantages in biomedical optics, data transmission, photomedicine, and detection of multi-scale phenomena. Recently, hydrogels have emerged as a promising candidate for interfacing photonics and bioengineering by combining their light-guiding properties with live tissue compatibility in optical, chemical, physiological, and mechanical dimensions. Herein, the latest progress over hydrogel photonics and its applications in guidance and manipulation of light is reviewed. Physics of guiding light through hydrogels and living tissues, and existing technical challenges in translating these tools into biomedical settings are discussed. A comprehensive and thorough overview of materials, fabrication protocols, and design architectures used in hydrogel photonics is provided. Finally, recent examples of applying structures such as hydrogel optical fibers, living photonic constructs, and their use as light-driven hydrogel robots, photomedicine tools, and organ-on-a-chip models are described. By providing a critical and selective evaluation of the field's status, this work sets a foundation for the next generation of hydrogel photonic research.

Photonic resonator interferometric scattering microscopy.

Interferometric scattering microscopy is increasingly employed in biomedical research owing to its extraordinary capability of detecting nano-objects individually through their intrinsic elastic scattering. To significantly improve the signal-to-noise ratio without increasing illumination intensity, we developed photonic resonator interferometric scattering microscopy (PRISM) in which a dielectric photonic crystal (PC) resonator is utilized as the sample substrate. The scattered light is amplified by the PC through resonant near-field enhancement, which then interferes with the <1% transmitted light to create a large intensity contrast. Importantly, the scattered photons assume the wavevectors delineated by PC's photonic band structure, resulting in the ability to utilize a non-immersion objective without significant loss at illumination density as low as 25 W cm-2. An analytical model of the scattering process is discussed, followed by demonstration of virus and protein detection. The results showcase the promise of nanophotonic surfaces in the development of resonance-enhanced interferometric microscopies.

Simulations and experimental demonstration of three different regimes of optofluidic manipulation.

It has been demonstrated that optically controlled microcurrents can be used to capture and move around a variety of microscopic objects ranging from cells and nanowires to whole live worms. Here, we present our findings on several new regimes of optofluidic manipulation that can be engineered using careful design of microcurrents. We theoretically optimize these regimes using COMSOL Multiphysics and present three sets of simulations and corresponding optofluidic experiments. In the first regime, we use local fluid heating to create a microcurrent with a symmetric toroid shape capturing particles in the center. In the second regime, the microcurrent shifts and tilts because external fluid flow is introduced into the microfluidic channel. In the third regime, the whole microfluidic channel is tilted, and the resulting microcurrent projects particles in a fan-like fashion. All three configurations provide interesting opportunities to manipulate small particles in fluid droplets and microfluidic channels.

Validation of an opto-electronic instrument for the measurement of execution velocity in squat exercise.

The purpose of this study was to analyse the reliability and validity of an opto-electronic sensor system (Velowin) for assessment of the bar-velocity in the deep squat exercise. Mean velocity, mean propulsive velocity and peak velocity generated in the deep squat exercise performed in the Smith machine bar were analysed compared to a linear velocity transducer considered as the gold standard. The study was conducted with a sample of 26 men with experience in resistance training. Six measurements were analysed for squat exercise in concentric phase using a progressive loading increase. Three consecutive repetitions were performed per load with a 3-4 min recovery between loads. Analysis of variance confirmed that there were no significant differences (p > 0.05) for the velocity variables between Velowin and T-Force for each of the loads. The reliability analysis showed high values of the intraclass correlation coefficient (ICC = 0.94-0.99), an 'almost perfect' Lin's concordance coefficient (CCC = 0.99) and a low coefficient of variation (CV <3.4%) for each of the loads and velocities. These results confirm the reliability and validity of the Velowin device for measuring the execution velocity in deep squat exercise.

Integrated Photonic System for Early Warning of Cyanobacterial Blooms in Aquaponics.

Cyanobacterial blooms produce hazardous toxins, deplete oxygen, and secrete compounds that confer undesirable organoleptic properties to water. To prevent bloom appearance, the World Health Organization has established an alert level between 500 and 2000 cells·mL-1, beyond the capabilities of most optical sensors detecting the cyanobacteria fluorescent pigments. Flow cytometry, cell culturing, and microscopy may reach these detection limits, but they involve both bulky and expensive laboratory equipment or long and tedious protocols. Thus, no current technology allows fast, sensitive, and in situ detection of cyanobacteria. Here, we present a simple, user-friendly, low-cost, and portable photonic system for in situ detection of low cyanobacterial concentrations in water samples. The system integrates high-performance preconcentration elements and optical components for fluorescence measurement of specific cyanobacterial pigments, that is, phycocyanin. Phycocyanin has demonstrated to be more selective to cyanobacteria than other pigments, such as chlorophyll-a, and to present an excellent linear correlation with bacterial concentration from 102 to 104 cell·mL-1 (R2 = 0.99). Additionally, the high performance of the preconcentration system leads to detection limits below 435 cells·mL-1 after 10 min in aquaponic water samples. Due to its simplicity, compactness, and sensitivity, we envision the current technology as a powerful tool for early warning and detection of low pathogen concentrations in water samples.

Matrix optics representation and imaging analysis of a light-field near-eye display.

Integral-imaging-based (InI-based) light-field near-eye display (LF-NED) is an effective way to relieve vergence-accommodation conflict (VAC) in applications of virtual reality (VR) and augmented reality (AR). Lenslet arrays are often used as spatial light modulator (SLM) in such systems. However, the conflict between refocusing on a virtual object point from the light-field image (LF image) and focusing on the image plane of the lenslets leads to degradation of the viewing effect. Thus, the light field (LF) cannot be accurately restored. In this study, we introduce matrix optics and build a parameterized model of a lenslet-array-based LF-NED with general applicability, based on which the imaging process is derived, and the performance of the system is analyzed. A lenslet-array-based LF-NED optical model is embodied in LightTools to verify the theoretical model. The simulations prove that the model we propose and the conclusions about it are consistent with the simulation results. Thus, the model can be used as the theoretical basis for evaluating the primary performance of an InI-based LF-NED system.

Image edge detection with a photonic spiking VCSEL-neuron.

We report both experimentally and in theory on the detection of edge features in digital images with an artificial optical spiking neuron based on a vertical-cavity surface-emitting laser (VCSEL). The latter delivers fast (< 100 ps) neuron-like optical spikes in response to optical inputs pre-processed using convolution techniques; hence representing image feature information with a spiking data output directly in the optical domain. The proposed technique is able to detect target edges of different directionalities in digital images by applying individual kernel operators and can achieve complete image edge detection using gradient magnitude. Importantly, the neuromorphic (brain-like) spiking edge detection of this work uses commercially sourced VCSELs exhibiting responses at sub-nanosecond rates (many orders of magnitude faster than biological neurons) and operating at the important telecom wavelength of 1300 nm; hence making our approach compatible with optical communication and data-centre technologies.

Photonic-chip assisted correlative light and electron microscopy.

Correlative light and electron microscopy (CLEM) unifies the versatility of light microscopy (LM) with the high resolution of electron microscopy (EM), allowing one to zoom into the complex organization of cells. Here, we introduce photonic chip assisted CLEM, enabling multi-modal total internal reflection fluorescence (TIRF) microscopy over large field of view and high precision localization of the target area of interest within EM. The photonic chips are used as a substrate to hold, to illuminate and to provide landmarking of the sample through specially designed grid-like numbering systems. Using this approach, we demonstrate its applicability for tracking the area of interest, imaging the three-dimensional (3D) structural organization of nano-sized morphological features on liver sinusoidal endothelial cells such as fenestrations (trans-cytoplasmic nanopores), and correlating specific endo-lysosomal compartments with its cargo protein upon endocytosis.