Direct Detection Device Research Articles

Non-enzymatic paper-based analytical device for direct potentiometric detection of urine creatinine.

A paper-based analytical device (PAD) with an integrated composite electrode has been designed and fabricated for non-enzymatic creatinine sensing. Reduced graphene oxide (rGO) was employed to modify the PAD so that it could function as a solid-contact transducer. A new macrocyclic pyrido-hexapeptide derivative was made and used as a special ionophore in the creatinine membrane sensor. The synthesized PAD showed a detection limit of 1.0µM (S/N = 3) and a potentiometric response towards creatinine throughout a log-linear range of 2.0µM-10mM (R2 = 0.9998). The sensor shows significant selectivity for a few related substances, including ephedrine, codeine, ketamine, caffeine, urea, urate, carbinoxamine, and dextromethorphan. It has been established that the testing method is appropriate for the direct potentiometric detection of creatinine in a variety of human urine sample types. When an indicating electrode and a reference electrode are put on the same flexible disposable, this lets applications with a small sample volume be done. For point-of-care creatinine measurement, the developedpaper-based analytical equipment is a good choice because it is affordable, easily accessible, and self-pumping (especially when combined with potentiometric detection).

MEPSi: A tool for simulating tomograms of membrane-embedded proteins

The throughput and fidelity of cryogenic cellular electron tomography (cryo-ET) is constantly increasing through advances in cryogenic electron microscope hardware, direct electron detection devices, and powerful image processing algorithms. However, the need for careful optimization of sample preparations and for access to expensive, high-end equipment, make cryo-ET a costly and time-consuming technique. Generally, only after the last step of the cryo-ET workflow, when reconstructed tomograms are available, it becomes clear whether the chosen imaging parameters were suitable for a specific type of sample in order to answer a specific biological question. Tools for a-priory assessment of the feasibility of samples to answer biological questions and how to optimize imaging parameters to do so would be a major advantage. Here we describe MEPSi (Membrane Embedded Protein Simulator), a simulation tool aimed at rapid and convenient evaluation and optimization of cryo-ET data acquisition parameters for studies of transmembrane proteins in their native environment. We demonstrate the utility of MEPSi by showing how to detangle the influence of different data collection parameters and different orientations in respect to tilt axis and electron beam for two examples: (1) simulated plasma membranes with embedded single-pass transmembrane αIIbβ3 integrin receptors and (2) simulated virus membranes with embedded SARS-CoV-2 spike proteins.

An electron counting algorithm improves imaging of proteins with low-acceleration-voltage cryo-electron microscope

Relative to the 300-kV accelerating field, electrons accelerated under lower voltages are potentially scattered more strongly. Lowering the accelerate voltage has been suggested to enhance the signal-to-noise ratio (SNR) of cryo-electron microscopy (cryo-EM) images of small-molecular-weight proteins (<100 kD). However, the detection efficient of current Direct Detection Devices (DDDs) and temporal coherence of cryo-EM decrease at lower voltage, leading to loss of SNR. Here, we present an electron counting algorithm to improve the detection of low-energy electrons. The counting algorithm increased the SNR of 120-kV and 200-kV cryo-EM image from a Falcon III camera by 8%, 20% at half the Nyquist frequency and 21%, 80% at Nyquist frequency, respectively, resulting in a considerable improvement in resolution of 3D reconstructions. Our results indicate that with further improved temporal coherence and a dedicated designed camera, a 120-kV cryo-electron microscope has potential to match the 300-kV microscope at imaging small proteins.

A point-of-care microfluidic channel-based device for rapid and direct detection of fibrinogen in whole blood

Hemorrhage is the leading cause of preventable death in civilian and battlefield traumatic injuries. Patients with severe traumatic hemorrhagic shock are more likely to be deficient in fibrinogen than those with other coagulation factors, and hypofibrinogenemia is an independent risk factor for mortality. Thus, rapid detection of fibrinogen levels is of great importance in these patients during damage control resuscitation. Plasma is used as an analyte in fibrinogen detection, which restricts the use of existing devices in emergencies. To meet the needs of on-site detection, we developed a point-of-care microfluidic channel-based device for direct measurement of fibrinogen concentration in whole blood. In our method, thrombin is dispersed on a reaction strip to initiate conversion of fibrinogen to fibrin. The permeability of the resulting blood clots depends on the fibrinogen level. A hydrophobic plastic protection flake between the reaction strip and a wicking strip is then removed to allow flow of unclotted blood. The rate of blood flow along the wicking strip was inversely related to the fibrinogen concentration. The whole process could be completed in as fast as 5 minutes for a whole blood sample size of 150 μL, and yielded accurate results ranging from 0 to 4 g L-1, which were unaffected by Ca2+, blood lipids, hematocrit, warfarin and tissue plasminogen activators (tPAs). Results using clinical whole blood samples were also highly consistent with those using an automatic coagulation analyzer, yielding a Pearson correlation coefficient of up to 0.919. This approach has potential for allowing rapid diagnosis of fibrinogen concentration in critically ill bleeding patients in different settings, thus helping to judge the suitability of fibrinogen replacement therapy (FRT) in cases of emergency bleeding and in patients at risk of thrombosis due to hyperfibrinogenemia.

Computational evaluation of sparse coding on off-axis electron holograms: comparison between charge-coupled device and direct-detection device cameras.

The effectiveness of sparse coding for image inpainting and denoising of off-axis electron holograms was examined computationally based on hologram simulations according to considerations of two types of electron detectors, namely charge-coupled device (CCD) and direct-detection device (DDD) cameras. In this simulation, we used a simple-phase object with a phase step such as a semiconductor p-n junction and assumed that the holograms recorded by the CCD camera include shot noise, dark-current noise and read-out noise, while those recorded by the DDD camera include only shot noise. Simulated holograms with various electron doses were sparsely coded. Even though interference fringes cannot be recognized in the simulated CCD and DDD holograms when subjected to electron doses (per pixel) equal to 1 and 0.01, respectively, both the corresponding sparse-coded holograms exhibit meaningful interference fringes. We demonstrate that a combination of the DDD camera and sparse coding reduces the requisite dose used to obtain holograms to values less than one-thousandth compared with the CCD camera without image postprocessing. This combination is expected to generate lower-dose and/or higher-speed electron holography.

Electrochemical Microsensor for Microfluidic Glyphosate Monitoring in Water Using MIP-Based Concentrators.

Glyphosate (GLY) is a broad-spectrum herbicide and is the most used pesticide worldwide. This vast usage has raised strong interest in the ecotoxicological impacts and human risks, with contamination of water being a major concern. Decentralized analytical techniques for water monitoring are of high importance. In this work, we present a small, low-cost, and time-effective electrochemical, chip-based microfluidic device for direct electrochemical detection of GLY downstream of a molecularly imprinted polymer (MIP) concentrator. We studied the electrochemical behavior of GLY and its metabolite aminomethylphosphonic acid (AMPA) using cyclic voltammetry with noble metal electrodes in acidic, neutral, and basic media. A chronoamperometric sensor protocol was developed for sensitive and selective GLY measurements on gold electrodes. The optimized protocol was transferred to a chip-based microsensor platform for online and real-time detection of GLY in a microfluidic setup. The results in the range from 0 to 50 μM GLY in 0.5 M H2SO4 show high linearity and a sensitivity of 10.3 ± 0.6 μA mm-2 mM-1 for the chip-based microfluidic platform. Successful recovery of GLY concentrated from untreated tap water and its precise detection from low volumes demonstrates the advantages of our system.

Easy Surface Functionalization and Bioconjugation of Peptides as Capture Agents of a Microfluidic Biosensing Platform for Multiplex Assay in Serum.

The development of assays for protein biomarkers in complex matrices is a demanding task that still needs implementation of new approaches. Antibodies as capture agents have been largely used in bioassays but their low stability, low-efficiency production, and cross-reactivity in multiplex approaches impairs their larger applications. Instead, synthetic peptides, even with higher stability and easily adapted amino acid sequences, still remain largely unexplored in this field. Here, we provide a proof-of-concept of a microfluidic device for direct detection of biomarker overexpression. The multichannel microfluidic polydimethylsiloxane (PDMS) device was first derivatized with PAA (poly(acrylic acid)) solution. CRP-1, VEGF-114, and ΦG6 peptides were preliminarily tested to respectively bind the biomarkers, C-reactive protein (CRP), vascular endothelial growth factor (VEGF), and tumor necrosis factor-alpha (TNF-α). Each PDMS microchannel was then respectively bioconjugated with a specific peptide (CRP-1, VEGF-114, or ΦG6) to specifically capture CRP, VEGF, and TNF-α. With such microdevices, a fluorescence bioassay has been set up with sensitivity in the nanomolar range, both in buffered solution and in human serum. The proposed multiplex assay worked with a low amount of sample (25 μL) and detected biomarker overexpression (above nM concentration), representing a noninvasive and inexpensive screening platform.

Elemental mapping of labelled biological specimens at intermediate energy loss in an energy-filtered TEM acquired using a direct detection device.

The technique of colour EM that was recently developed enabled localisation of specific macromolecules/proteins of interest by the targeted deposition of diaminobenzidine (DAB) conjugated to lanthanide chelates. By acquiring lanthanide elemental maps by energy‐filtered transmission electron microscopy (EFTEM) and overlaying them in pseudo‐colour over the conventional greyscale TEM image, a colour EM image is generated. This provides a powerful tool for visualising subcellular component/s, by the ability to clearly distinguish them from the general staining of the endogenous cellular material. Previously, the lanthanide elemental maps were acquired at the high‐loss M4,5 edge (excitation of 3d electrons), where the characteristic signal is extremely low and required considerably long exposures. In this paper, we explore the possibility of acquiring the elemental maps of lanthanides at their N4,5 edge (excitation of 4d electrons), which occurring at a much lower energy‐loss regime, thereby contains significantly greater total characteristic signal owing to the higher inelastic scattering cross‐sections at the N4,5 edge. Acquiring EFTEM lanthanide elemental maps at the N4,5 edge instead of the M4,5 edge, provides ∼4× increase in signal‐to‐noise and ∼2× increase in resolution. However, the interpretation of the lanthanide maps acquired at the N4,5 edge by the traditional 3‐window method, is complicated due to the broad shape of the edge profile and the lower signal‐above‐background ratio. Most of these problems can be circumvented by the acquisition of elemental maps with the more sophisticated technique of EFTEM Spectrum Imaging (EFTEM SI). Here, we also report the chemical synthesis of novel second‐generation DAB lanthanide metal chelate conjugates that contain 2 lanthanide ions per DAB molecule in comparison with 0.5 lanthanide ion per DAB in the first generation. Thereby, fourfold more Ln3+ per oxidised DAB would be deposited providing significant amplification of signal. This paper applies the colour EM technique at the intermediate‐loss energy‐loss regime to three different cellular targets, namely using mitochondrial matrix‐directed APEX2, histone H2B‐Nucleosome and EdU‐DNA. All the examples shown in the paper are single colour EM images only.

Advances in cryo-EM and ED with a cold-field emission beam and energy filtration -Refinements of the CRYO ARM 300 system in RIKEN SPring-8 center.

We have designed and evaluated a cryo-electron microscopy (cryo-EM) system for higher-resolution single particle analysis and high-precision electron 3D crystallography. The system comprises a JEOL CRYO ARM 300 electron microscope-the first machine of this model-and a direct detection device camera, a scintillator-coupled camera, GPU clusters connected with a camera control computer and software for automated-data collection and efficient and accurate operation. The microscope provides parallel illumination of a highly coherent 300-kV electron beam to a sample from a cold-field emission gun and filters out energy-loss electrons through the sample with an in-column energy filter. The gun and filter are highly effective in improving imaging and diffraction, respectively, and have provided high quality data since July 2018. We here report on the characteristics of the cryo-EM system, updates, our progress and future plan for running such cryo-EM machines in RIKEN SPring-8 Center.

Phase detection limits in off-axis electron holography from pixelated detectors: gain variations, geometric distortion and failure of reference-hologram correction.

We investigate the effect that recording off-axis electron holograms on pixelated detectors, such as charge-coupled devices (CCD) and direct-detection devices (DDD), can have on measured amplitudes and phases. Theory will be developed for the case of perfectly uniform interference fringes illuminating an imperfect detector with gain variations and pixel displacements. We will show that both these types of defect produce a systematic noise in the phase images that depends on the position of the holographic fringes with respect to the detector. Subtracting a reference hologram from the object hologram will therefore not remove the phase noise if the initial phases of the two holograms do not coincide exactly. Another finding is that pi-shifted holograms are much less affected by gain variations but show no improvement concerning geometric distortions. The resulting phase errors will be estimated and simulations presented that confirm the theoretical developments.

A Nanostructured Gold/Graphene Microfluidic Device for Direct and Plasmonic-Assisted Impedimetric Detection of Bacteria.

Hierarchical 3D gold nano-/microislands (NMIs) are favorably structured for direct and probe-free capture of bacteria in optical and electrochemical sensors. Moreover, their unique plasmonic properties make them a suitable candidate for plasmonic-assisted electrochemical sensors, yet the charge transfer needs to be improved. In the present study, we propose a novel plasmonic-assisted electrochemical impedimetric detection platform based on hybrid structures of 3D gold NMIs and graphene (Gr) nanosheets for probe-free capture and label-free detection of bacteria. The inclusion of Gr nanosheets significantly improves the charge transfer, addressing the central issue of using 3D gold NMIs. Notably, the 3D gold NMIs/Gr detection platform successfully distinguishes between various types of bacteria including Escherichia coli (E. coli) K12, Pseudomonas putida (P. putida), and Staphylococcus epidermidis (S. epidermidis) when electrochemical impedance spectroscopy is applied under visible light. We show that distinguishable and label-free impedimetric detection is due to dissimilar electron charge transfer caused by various sizes, morphologies, and compositions of the cells. In addition, the finite-difference time-domain (FDTD) simulation of the electric field indicates the intensity of charge distribution at the edge of the NMI structures. Furthermore, the wettability studies demonstrated that contact angle is a characteristic feature of each type of captured bacteria on the 3D gold NMIs, which strongly depends on the shape, morphology, and size of the cells. Ultimately, exposing the platform to various dilutions of the three bacteria strains revealed the ability to detect dilutions as low as ∼20 CFU/mL in a wide linear range of detection of 2 × 101-105, 2 × 101-104, and 1 × 102-1 × 105 CFU/mL for E. coli, P. putida, and S. epidermidis, respectively. The proposed hybrid structure of 3D gold NMIs and Gr, combined by novel plasmonic and conventional impedance spectroscopy techniques, opens interesting avenues in ultrasensitive label-free detection of bacteria with low cost and high stability.

Compact Grating-Coupled Biosensor for the Analysis of Thrombin

A compact optical biosensor for direct detection of thrombin in human blood plasma (HBP) is reported. This biosensor platform is based on wavelength spectroscopy of diffraction-coupled surface plasmons on a chip with a periodically corrugated gold film that carries an antifouling thin polymer layer consisting of poly[(N-(2-hydroxypropyl)methacrylamide)-co-(carboxybetaine methacrylamide)] (poly(HPMA-co-CBMAA)) brushes. This surface architecture provides superior resistance to nonspecific and irreversible adsorption of abundant compounds in the analyzed HBP samples in comparison to standard surface modifications. The carboxylate groups along the polymer brushes were exploited for the covalent immobilization of aptamer ligands. These ligands were selected to specifically capture the target thrombin analyte from the analyzed HBP sample in a way that does not activate the coagulatory process at the biosensor surface with poly(HPMA-co-CBMAA) brushes. Direct label-free analysis of thrombin in the medically relevant concentration range (1-20 nM) is demonstrated without the need for diluting the HBP samples or using additional steps for signal enhancement. The reported platform constitutes the first step toward a portable and sensitive point-of-care device for direct detection of thrombin in human blood.

36. Optimization of the acquisition threshold of Photon Counting Detectors (PCDs) used in X-ray medical imaging

Purpose PCDs with energy discrimination led to new possibilities of improving the quality of X-ray medical images. PCDs are direct detection devices in which the charge delivered by a single interacting photon is directly collected and discriminated at single pixels level. The limit of high densely pixelated PCDs is the charge sharing which can lead to multiple counts from a single interaction degrading the spatial resolution of the system. Increasing the discrimination threshold, multiple counts can be partially or totally removed from images improving the spatial resolution at the cost of a reduction of the detection area of single pixels with a consequent loss of statistics. This work aims to show a simple method to find the optimal threshold which gives the best trade-off between spatial resolution and statistics. Methods The acquisitions have been performed at the SYRMEP beamline (Elettra Trieste) using the tomographic experimental set-up of INFN project SYnchrotron Radiation MAmmography (SYRMA-3D). SYRMA-3D uses ‘Pixirad-8’, a PCD with CdTe sensor and a pixel size of 60 μm. For this study, we used a monochromatic beam of 38 keV acquiring a set of images of an edge and a set of images of the beam with different detection thresholds. The two sets of images have been used respectively for the measurement of the Modulation transfer function (MTF) and the noise power spectrum (NPS). Combining both NPS and MTF the Number of Equivalent Quanta (NEQ) can be determined. The NEQ summarizes both informations about spatial resolution and noise and is an useful metrics for the global image quality assessment. Results The comparison between NEQ curves shows that lower thresholds give better NEQ response. Conclusions The increase of the discrimination threshold improves the spatial resolution but worsens the noise. The global image quality, measured using NEQ, is higher for lower thresholds.

Three-dimensional electron microscopy techniques for unravelling mitochondrial dysfunction in heart failure and identification of new pharmacological targets.

A hallmark of heart failure is mitochondrial dysfunction leading to a bioenergetics imbalance in the myocardium. Consequently, there is much interest in targeting mitochondrial abnormalities to attenuate the pathogenesis of heart failure. This review discusses (i) how electron microscopy (EM) techniques have been fundamental for the current understanding of mitochondrial structure–function, (ii) the paradigm shift in resolutions now achievable by 3‐D EM techniques due to the introduction of direct detection devices and phase plate technology, and (iii) the application of EM for unravelling mitochondrial pathological remodelling in heart failure. We further consider the tremendous potential of multi‐scale EM techniques for the development of therapeutics, structure‐based ligand design and for delineating how a drug elicits nanostructural effects at the molecular, organelle and cellular levels. In conclusion, 3‐D EM techniques have entered a new era of structural biology and are poised to play a pivotal role in discovering new therapies targeting mitochondria for treating heart failure.Linked ArticlesThis article is part of a themed section on Mitochondrial Pharmacology: Featured Mechanisms and Approaches for Therapy Translation. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v176.22/issuetoc

Wax-printed paper-based device for direct electrochemical detection of 3-nitrotyrosine

An overview of the last trends in lab-on-chip technology highlights the continuous development of biosensing approaches in terms of simplicity, low-cost, and quick response, with a higher sensitivity and selectivity. In this context, electrochemical sensors arrived to overcome the need to develop small, accurate, portable and compact sensing devices, compatible with routine use in loco analysis, especially in point-of-care (POC) assessment.Herein, we have presented the first paper-based electrochemical sensor towards the detection of 3-nitrotyrosine (3-NT), a relevant oxidative stress biomarker. This innovative sensor was designed by, first, modifying paper to become a hydrophobic support and, second applying carbon and silver conductive inks to generate a three electrode-system on a small spot. An intensive voltammetric investigation of the paper-based screen-printed electrodes (SPEs), including the study of different redox-probes, scan-rate values, probe concentrations and supporting electrolyte solutions, confirmed their suitable and reliable electrochemical performance. Square wave voltammetry (SWV) was employed to assess the sensitivity of the proposed sensor at various concentrations of 3-NT, ranging from 500 nM to 1 mM, enabling a low detection limit (LOD) of 49.2 nM. The interference of co-existing species was also tested and good selectivity against 3-NT was accomplished. Overall, this flexible paper-based platform constitutes a promising sensor material towards a quick, sensitive, cost-effective, sustainable and reproducible detection of 3-NT biomarker.

MRCZ - A file format for cryo-TEM data with fast compression.

The introduction of fast CMOS detectors is moving the field of transmission electron microscopy into the computer science field of big data. Automated data pipelines control the instrument and initial processing steps which imposes more onerous data transfer and archiving requirements. Here we conduct a technical demonstration whereby storage and read/write times are improved 10× at a dose rate of 1 e-/pix/frame for data from a Gatan K2 direct-detection device by combination of integer decimation and lossless compression. The example project is hosted at github.com/em-MRCZ and released under the BSD license.

Using Tomoauto: A Protocol for High-throughput Automated Cryo-electron Tomography.

Cryo-electron tomography (Cryo-ET) is a powerful three-dimensional (3-D) imaging technique for visualizing macromolecular complexes in their native context at a molecular level. The technique involves initially preserving the sample in its native state by rapidly freezing the specimen in vitreous ice, then collecting a series of micrographs from different angles at high magnification, and finally computationally reconstructing a 3-D density map. The frozen-hydrated specimen is extremely sensitive to the electron beam and so micrographs are collected at very low electron doses to limit the radiation damage. As a result, the raw cryo-tomogram has a very low signal to noise ratio characterized by an intrinsically noisy image. To better visualize subjects of interest, conventional imaging analysis and sub-tomogram averaging in which sub-tomograms of the subject are extracted from the initial tomogram and aligned and averaged are utilized to improve both contrast and resolution. Large datasets of tilt-series are essential to understanding and resolving the complexes at different states, conditions, or mutations as well as obtaining a large enough collection of sub-tomograms for averaging and classification. Collecting and processing this data can be a major obstacle preventing further analysis. Here we describe a high-throughput cryo-ET protocol based on a computer-controlled 300kV cryo-electron microscope, a direct detection device (DDD) camera and a highly effective, semi-automated image-processing pipeline software wrapper library tomoauto developed in-house. This protocol has been effectively utilized to visualize the intact type III secretion system (T3SS) in Shigella flexneri minicells. It can be applicable to any project suitable for cryo-ET.

Two promising future developments of cryo-EM: capturing short-lived states and mapping a continuum of states of a macromolecule.

The capabilities and application range of cryogenic electron microscopy (cryo-EM) method have expanded vastly in the last two years, thanks to the advances provided by direct detection devices and computational classification tools. We take this review as an opportunity to sketch out promising developments of cryo-EM in two important directions: (i) imaging of short-lived states (10-1000 ms) of biological molecules by using time-resolved cryo-EM, particularly the mixing-spraying method and (ii) recovering an entire continuum of coexisting states from the same sample by employing a computational technique called manifold embedding. It is tempting to think of combining these two methods, to elucidate the way the states of a molecular machine such as the ribosome branch and unfold. This idea awaits further developments of both methods, particularly by increasing the data yield of the time-resolved cryo-EM method and by developing the manifold embedding technique into a user-friendly workbench.

The influence of frame alignment with dose compensation on the quality of single particle reconstructions.

As direct electron detection devices in cryo-electron microscopy become ubiquitous, the field is now ripe for new developments in image analysis techniques that take advantage of their increased SNR coupled with their high-throughput frame collection abilities. In approaching atomic resolution of native-like biomolecules, the accurate extraction of structural locations and orientations of side-chains from frames depends not only on the electron dose that a sample receives but also on the ability to accurately estimate the CTF. Here we use a new 2.8Å resolution structure of a recombinant gene therapy virus, AAV-DJ with Arixtra, imaged on an FEI Titan Krios with a DE-20 direct electron detector to probe new metrics including relative side-chain density and ResLog analysis for optimizing the compensation of electron beam damage and to characterize the factors that are limiting the resolution of the reconstruction. The influence of dose compensation on the accuracy of CTF estimation and particle classifiability are also presented. We show that rigorous dose compensation allows for better particle classifiability and greater recovery of structural information from negatively charged, electron-sensitive side-chains, resulting in a more accurate macromolecular model.

Development of a fiber daylighting system based on the parallel mechanism and direct focus detection

In this paper, a fiber optic daylighting system based on the parallel mechanism is designed and fabricated. This system is composed of 49 concentrating cells, which are arranged in a 7×7 array. The array consists of 48 sunlight-collecting cells and a sun position sensor using direct focus detection. A linear actuator, which is composed of two stepper motors and two roller screws, is used to drive the concentrating array to track the sun with the movement in a plane parallel to the plane containing the array of fiber optic collecting aperture. The solid angle of the sun tracking angle range for the concentrating cells is 4π/3 sr (covering 2/3 of the sky), satisfying the demand for 8h of tracking. This system is suitable for building integration because it has a compact and flat shape. The sun position sensor consists of photodiodes and a fiber array in the shape of a cross. The sun position sensor can directly measure the high flux density of the focus (2500 suns). A series of tests were performed using a lux meter and spectrometer to investigate the photometric characteristics of the system for a lightless underpass of 8.6m×4.2m×2.3m. The experiments show that the transfer efficiency of this system can reach 25% (with a 10-m long fiber). The luminous efficacy can reach 250lum/W, which is two times than that of natural light because the fiber can filter out infrared light. The experiments also verified the feasibility of the daylighting system using the parallel mechanism and a direct detection device for highly concentrated light.