Amplification System Research Articles

Tissue-Specific RNA Localization in the Uterus During Implantation, Decidualization and Placentation: Technical Nuances of Various Labeling Approaches.

In situ hybridization (ISH) is a sensitive method used to localize a specific sequence of DNA or RNA in biological samples, including cells, tissue sections or whole organs. RNA ISH can be used to determine spatial gene expression using a single-stranded probe with a reverse-complementary sequence. Cell-specific gene expression has been studied using mRNA and protein levels. Signals produced by RNA probes are usually more specific than those produced by antibodies in immunostaining. Currently, ISH is the most widely used method to localize mRNA molecules. Traditionally, probes were labeled with radioactive isotopes, but the cumbersome procedures and potential health risk limit their acceptance. Recently, probes labeled with nonradioactive materials including digoxigenin, biotin and various fluorophores have been developed. The tyramide signal amplification system further enhances the sensitivity of detection. These methods have been applied in numerous studies in various tissues including reproductive organs. This article details three methods of RNA in situ hybridization: radioactive in situ hybridization, digoxigenin in situ hybridization, and digoxigenin-tyramide signal amplification fluorescein in situ hybridization. The pros and cons of each protocol are discussed. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Radioactive in situ hybridization (radioactive-ISH) Basic Protocol 2: Digoxigenin in situ hybridization (DIG-ISH) Basic Protocol 3: Digoxigenin-tyramide signal amplification fluorescein in situ hybridization (DIG-TSA-FISH).

A triple-cycle amplification biosensor for colorimetric detection of mutant PIK3CAE545K based on cascade-driven DNA walker and branched hybridization strand reaction

Circulating tumor DNA (ctDNA) is an ideal candidate for liquid biopsy biomarkers. Therefore, detecting a low abundance of ctDNA is essential for early cancer diagnosis. Here, we developed a novel triple circulation amplification system integrating entropy and enzyme cascade-driven three-dimensional (3D) DNA walker and branched hybridization strand reaction (B-HCR) for ultrasensitive detection of breast cancer-related ctDNA. In this study, the 3D DNA walker was constructed by inner track probes (NH) and complex S on a microsphere. Once the DNA walker was triggered by the target, the strand replacement reaction ran first and kept circulating to rapidly displace the DNA walker containing 8–17 DNAzyme. Secondly, the DNA walker could repeatedly cleave NH autonomously along the inner track, generating numerous initiators, and then promoting B-HCR to activate the third cycle. Subsequently, the split G-rich fragments were brought in close to form the G-quadruplex/hemin DNAzyme by adding hemin, with the addition of H2O2 and ABTS, the target could be observed. Benefiting from triplex cycles, the PIK3CAE545K mutation detection possesses a good linear range from 1-103 fM, and the limit of detection was 0.65 fM. Due to the low cost and high sensitivity, the proposed strategy has great potential in early diagnosis of breast cancer.

Detection of Hazelnut and Almond Adulteration in Olive Oil: An Approach by qPCR.

Virgin olive oil (VOO), characterized by its unique aroma, flavor, and health benefits, is subject to adulteration with the addition of oils obtained from other edible species. The consumption of adulterated olive oil with nut species, such as hazelnut or almond, leads to health and safety issues for consumers, due to their high allergenic potential. To detect almond and hazelnut in olive oil, several amplification systems have been analyzed by qPCR assay with a SYBR Green post-PCR melting curve analysis. The systems selected were Cora1F2/R2 and Madl, targeting the genes coding the allergenic protein Cor a 1 (hazelnut) and Pru av 1 (almond), respectively. These primers revealed adequate specificity for each of the targeted species. In addition, the result obtained demonstrated that this methodology can be used to detect olive oil adulteration with up to 5% of hazelnut or almond oil by a single qPCR assay, and with a level as low as 2.5% by a nested-qPCR assay. Thus, the present research has shown that the SYBR-based qPCR assay can be a rapid, precise, and accurate method to detect adulteration in olive oil.

Highly efficient near-infrared electrochemiluminescence resonance energy transfer system for biosensing: Nonmetallic plasmon Mediated well-matched energy donor-acceptor pair

Herein, a well-matched energy donor-acceptor pair strategy was designed to construct highly efficient NIR ECL-RET system. In detail, an ECL amplification system consisting of SnS2 quantum dots decorated Ti3C2 MXene nanocomposites (SnS2 QDs-Ti3C2) as the energy donor were developed via a one-pot method, and the nanocomposites exhibited highly efficient NIR ECL emission due to the surface-defect effect generated by the oxygen-containing functional groups in MXene. Nonmetallic plasmon hydrated defective tungsten oxide nanosheets (dWO3•H2O) were utilized as energy acceptors because of its strong surface plasmon resonance effect in Vis-NIR absorption range. Compared with non-defective tungsten oxide hydrate nanosheets (WO3•H2O), the overlapping area between ECL spectrum of SnS2 QDs-Ti3C2 and UV–vis spectrum of dWO3•H2O was increased by 2.1 times, and the results showed that more efficient quenching effect was obtained. As a proof of concept, tetracycline (TCN) aptamer and its complementary chain were served as a bridge to connect the energy donor and acceptor, achieving the successful construction of NIR ECL-RET aptasensor. The as-fabricated ECL sensing platform exhibited a low detection limit of 6.2 fM (S/N = 3) within a wide linear range from 10 fM to 10 μM. Besides, the NIR ECL-RET aptasensor also showed excellent stability, reproducibility and selectivity, providing a promising tool to detect TCN in real samples. This strategy offered a universal and effective method in constructing highly efficient NIR ECL-RET system for developing rapid, sensitive and accurate biological detection platform.

Finite-amplitude nonlinear waves in inertial amplification metamaterials: Theoretical and numerical analyses

The motivation of this study is to understand the unique characteristics of finite-amplitude elastic wave propagation in an inertial amplification (IA) system coupled with a Duffing resonator and leverage the unique opportunities offered by the IA mechanism and nonlinearities to achieve the modulation of elastic waves. Based on the assumption of the perturbation method, the effective range of the wave amplitude was investigated, which is usually ignored. Meanwhile, the obvious phenomenon of bandgap transition (between the single resonance bandgap and hybrid bandgap) is observed with varying IA levels. In particular, we demonstrated a new phenomenon of negative wave propagation and an obvious standing-wave mode in this system. Here, an analytical investigation of the amplitude-dependent and IA-dependent nonlinear dispersion is conducted based on the perturbation approach. Numerical simulations using the time-domain finite element method and Gaussian pulse excitation were performed to validate the nonlinear dispersion and negative motion of the wave packet. The research results can be used to tune wave propagation in metamaterials and provide ideas for the design of metamaterials, which can be used in practical circumstances for different energy transfer needs.

Calibration-free disposable electrochemical sensor with co-facing electrodes: Theory and characterisation with fixed and changing mediator concentration

The use of electrochemical sensors containing oxidoreductase enzymes or redox-active substrates has become an area of particular interest due to its wide potential applications. This work presents the construction, theory and characterisation of a calibration-free disposable electrochemical sensor with co-facing electrodes. Each co-facing electrode is used as an anode or cathode to achieve high sensitivity based on a reversible redox mediator-recycling signal amplification system. Furthermore, this system can simultaneously report on the concentration and diffusion coefficient of the limiting electrochemical mediator. Supporting experimental evidence is provided in systems where the viscosity of the solution is systematically varied. The theory for a constant limiting electrochemical mediator concentration is presented and extended to treat the case where the mentioned concentration increases linearly with time. The theory is applied to three specific cases to validate the developed theory. The sensor presented in this paper may be applied to developing novel low-volume and high-sensitivity disposable, point-of-care electrochemical biosensors.

Messenger Materials Moving Forward: The Role of Functional Polymer Architectures as Enablers for Dynamic Nano‐to‐Macro Messaging

AbstractIdentifying changes in the nanoscopic domain is a key challenge in the physicochemical sciences, where great interest is on sensing complex processes that involve cellular biochemical reactions, chemical heterogeneities, contact forces, and other interfacial phenomena. This has stimulated the development of diverse materials that allow subtle nanoscopic environments to be "seen". The challenge in the nano‐domain has always been the ability to sense changes on the minute scale and rapidly transduce the information out for macroscopical observation. Ideally, materials should inform when processes are occurring. Recently, new systems that leverage established concepts with fluorescence‐ and plasmonic‐based sensing have been devised, which has reinvigorated the domain, where functional polymers coupled in specific architectures to transducing motifs allow for a new basis of messenger materials to be realized. The key aspect in this regard is that the polymers allow for sensing to be achieved only when they are carefully coupled to the amplification system. In this perspective, the role of specific functional polymer architectures for the realization of nano‐to‐macro sensing of subtle nano‐messengers is discussed and where the exciting field of messenger materials is seen moving forward is pointed out.

Fluidic Spatial-Confinement Scaffold Affords a Multicomponent DNA Reaction with Improved Efficiency and Accelerated Kinetics.

Enzyme-free nucleic acid amplification reactions with the capability of signal catalytic amplification have been widely used in biosensors. However, these multicomponent, multistep nucleic acid amplification systems often suffer from low reaction efficiency and kinetics. Herein, inspired by the natural cell membrane system, we utilized the red blood cell membrane as a fluidic spatial-confinement scaffold to construct a novel accelerated reaction platform. By simply modifying with cholesterol, DNA components can be efficiently integrated into the red blood cell membrane through hydrophobic interactions, which greatly increases the local concentration of DNA strands. Moreover, the fluidity of the erythrocyte membrane improves the collision efficiency of DNA components in the amplification system. Based on the increased local concentration and improved collision efficiency, the fluidic spatial-confinement scaffold significantly improved the reaction efficiency and kinetics. Taking catalytic hairpin assembly (CHA) as a model reaction, an RBC-CHA probe based on the erythrocyte membrane platform enables a more sensitive detection of miR-21 with a sensitivity that is 2 orders of magnitude higher than the free CHA probe and a fast reaction rate (about 3.3-fold). The proposed strategy provides a new idea for the construction of a novel spatial-confinement accelerated DNA reaction platform.

Development of a colloidal gold immunochromatographic strip with enhanced signal for the detection of bovine parvovirus.

Bovine parvovirus (BPV) is a pathogen responsible for respiratory and digestive tract symptoms in calves and abortion and stillbirth in pregnant cows. In this study, we developed a colloidal gold immunochromatographic (GICG) strip with an enhanced signal for detecting BPV according to the double-antibody sandwich principle and an enzyme-based signal amplification system to amplify the signal. This system utilizes horseradish peroxidase reacting with a substrate solution containing 3,3',5,5'-tetramethylbenzidine and dextran sulfate to obtain insoluble blue products on the test and control lines. We optimized different reaction conditions, including the amount of monoclonal antibodies (mAbs), pH of the colloidal gold solution, coating solution, blocking solution, sample pad treatment solution, antibody concentration in the control line, and antibody concentration in the detection line. The sensitivity of the signal-enhanced GICG strip showed that the minimum amount for detecting BPV was 102 TCID50, 10 times higher than that of the traditional GICG strip. The results of the specificity test showed that the signal-enhanced GICG strip had no cross-reactivity with BRV, BVDV, or BRSV. The results of the repeatability test showed that the coefficient of variation between and within batches was less than 5%, showing good repeatability. Moreover, for validation, PCR and the signal-enhanced GICG strip were used to detect 280 clinical bovine fecal samples. The concordance rate compared with PCR was 99.29%. Hence, the developed strip exhibited high sensitivity and specificity for the detection of BPV. Therefore, this strip could be a rapid, convenient, and effective method for the diagnosis of BPV infection in the field.

A Systematic Comparison of High-End and Low-Cost EEG Amplifiers for Concealed, Around-the-Ear EEG Recordings.

Wearable electroencephalography (EEG) has the potential to improve everyday life through brain-computer interfaces (BCI) for applications such as sleep improvement, adaptive hearing aids, or thought-based digital device control. To make these innovations more practical for everyday use, researchers are looking to miniaturized, concealed EEG systems that can still collect neural activity precisely. For example, researchers are using flexible EEG electrode arrays that can be attached around the ear (cEEGrids) to study neural activations in everyday life situations. However, the use of such concealed EEG approaches is limited by measurement challenges such as reduced signal amplitudes and high recording system costs. In this article, we compare the performance of a lower-cost open-source amplification system, the OpenBCI Cyton+Daisy boards, with a benchmark amplifier, the MBrainTrain Smarting Mobi. Our results show that the OpenBCI system is a viable alternative for concealed EEG research, with highly similar noise performance, but slightly lower timing precision. This system can be a great option for researchers with a smaller budget and can, therefore, contribute significantly to advancing concealed EEG research.

The effect of sound field amplification systems on vocal demand response in teachers during lessons

Purpose Sound pressure level (SPL), fundamental frequency (F0), time dose (Dt), cycle dose (Dc), and distance dose (Dd) are components affecting a vocal demand response. The study aimed to investigate the impact of sound field amplification systems (SFAS) on teachers’ vocal parameters and determine the user comfort of SFAS use in teachers. Methods Twenty female teachers underwent long-term voice dosimetry with Vocal Holter Med (PR.O.Voice Srl) during everyday teaching activities. The SFAS PentaClassRuna (Certes) was installed in classrooms. Voice dosimetry was provided in two different acoustical conditions—without the use of SFAS (one to two days) and with SFAS (one to three days). Teachers underwent voice acoustic and laryngoscopic evaluation before voice dosimetry. Two teachers’ groups were formed: teachers with and without vocal nodules. The visual analogue scale measured the user comfort of SFAS. Results There were no significant differences in vocal parameters and vocal doses between teachers with and without vocal nodules. Voice amplification significantly decreased average F 0(−4.4 Hz), Dt%(−3.1%), Dc (−0.4 kcycles), and Dd (−1.3 m) in teachers without vocal nodules and F 0 (−8.9 Hz) in teachers with vocal nodules. Vocal doses (Dt%, Dc, Dd) significantly decreased in classrooms with longer reverberation time. The user comfort of the SFAS during the lessons was high in both teachers groups. Conclusions SFAS was a mediator between the classroom’s environment and the teacher’s vocal demand response; it changed teachers’ voice production parameters and reduced vocal demand response to satisfy the communication requirements. In addition, voice amplification was more beneficial for teachers without vocal fold lesions.

Molecular quantification for differentiation of fresh and dried Jinqian Baihua She

Freshly-used crude drugs have unique functions and advantages in TCM practice of treating diseases. Jinlong Capsule is a patent traditional Chinese medicine product effective for treatment of hepatocarcinoma, and fresh Jinqian Baihua She (JBS, the body of juvenile Bungarus multicinctus) is one of its important ingredients. The emergence of counterfeit fresh JBS, often identified as dried JBS with almost identical appearance, poses a difficult problem in the quality control of the product. Herein we report a molecular quantification-based method for differentiation of fresh and dried JBS by determining the copy number of a specific DNA marker in the samples. Using species-specific primers and TaqMan probes, we established a real-time quantitative PCR system for amplification of a fragment in the 658-bp cytochrome oxidase subunit I (COI) region from JBS specimens. The amplicon copy number in the muscle tissues ranged from 1.14 × 107 to 4.83 × 107 copies/mg in fresh JBS samples, as compared with 1.13 × 105−8.91 × 106 copies/mg in dried JBS samples. Based upon Fisher discriminant analysis, we used 1.27 × 107 copies/mg as the cut-off value for differentiating fresh and dried JBS, which was validated in the single-blinded validation test of fresh and dried JBS samples. This qPCR system may provide an efficient means for accurate identification of fresh JBS to improve the quality control of the medicinal product.

Negative stiffness-enhanced seismic damping technology for an over-track complex with concrete-encased steel columns

The resilient transit-oriented model stimulates the necessity of seismic performance enhancement or retrofitting of over-track complexes with concrete-encased steel slender supporting columns. This study proposes a novel low-damping-ratio-based vibration control approach and easy-to-use design for over-track complexes by flexibly installing the negative stiffness amplification system (NSAS) at the multistory of the over-track building and lower podium. Moreover, a multilocation-oriented design strategy and fitted formulae are developed for the NSAS-damped complex with enhanced energy-dissipation efficiency and an adjustable structural modal shape. By utilizing the negative stiffness device, dashpot, and tuning spring, the NSAS is theoretically constructed, and a simplified model of the NSAS-damped over-track complex is established. Stochastic response analysis and parametric investigation are performed to quantify the benefit of the NSAS-damped story over the conventional control method. Then, easy-to-use design formulae are provided in the initial design and parameter modification stages to realize the simultaneous control of the upper and lower structures. Based on a typical over-track complex with concrete-encased steel slender supporting columns, design cases are analyzed for the NSAS and conventional viscous dampers to verify the applicability of the proposed NSAS and design. The results indicate that the target-story located NSASs contribute a higher-efficiency approach to realize the significant performance improvement or retrofit of the over-track complexes without perturbing the daily function. Particularly, NSASs tailored for distinct stories in the over-track complex effectively satisfy the modification demand for structural modal shape and enhanced energy dissipation efficiency, which cannot be realized by conventional viscous dampers or the existing design of NSASs in normal buildings. In addition, the NSAS-damped story demonstrates a released stiffness abruptness and robust seismic performances against various earthquakes.

Rational Design of Cascade DNA System for Signal Amplification.

System leakage critically confines the development of cascade DNA systems that need to be implemented in a strict order-by-order manner. In principle, ternary DNA reactants, composed of three single-strand DNA (ssDNA) with a strict equimolar ratio (1:1:1), have been indispensable for successfully cascading upstream entropy-driven DNA circuit (EDC) with downstream circuits, and system leakage will occur with any unbalance of the molar ratio. In this work, we proposed "splitting-reconstruction" and "protection-release" strategies on the potential downstream circuit initiator derived from upstream EDC to guide the construction of EDC-involved cascade systems independent of system leakage derived from unpurified reactants. Both the reconstructed and released downstream circuit initiators were in compliance with the principle of the cascade AND logic gate. Using these two strategies, two cascade systems─EDC2-4WJ-TMSDR and EDC3-HCR─were developed to carry out the designed order, which did not require that the ratio of 1:1:1 be maintained. Furthermore, the inherent property of the upstream EDC could transfer into the downstream circuit, endowing the developed cascade systems with a more powerful signal amplification ability for the sensitive detection of the corresponding initiator strand. These two strategies may provide new insights into the process of constructing EDC-like circuit-involved high-order DNA networks.

Research on A Fast Recognition System for Abnormal Harmonic Amplification Caused by Centralized Connection of Multiple Wind Power Plants

This paper introduces the abnormal harmonic current amplification phenomenon that occurred in the electric transmission lines, which is caused by the centralized join-up of multiple wind farm plants, by collecting and analysing the voltage, current, and active power indicators data of harmonic amplification phenomenon, to realize the classification and statistics of abnormal indexes variation trend during the harmonic amplification phenomenon. In this paper, the abnormal indexes are classified by different working conditions, such as normal operating state and harmonic amplification phenomenon, thus the recognition index of harmonic amplification is compared and summarized. After screening the most obvious changed part of the index under two kinds of working conditions, nine feature indexes are chosen to be embraced into the recognizing system. Finally, this method is used to analyse the harmonic amplification events in other moments of transmission lines, and all the harmonic amplification phenomena are identified successfully, which proves the feasibility and accuracy of this method.

CRISPR screening for cytotoxic granule degranulation pathway in CD8 T cell

Abstract Degranulation of perforin-containing cytotoxic granules is important for NK cells and CD8 cytotoxic T lymphocytes (CTLs) to kill infected cells or tumors. Defects in this pathway cause the primary form of hemophagocytic lymphohistiocytosis (HLH), which is known as familial HLH (FHL) syndrome. Most of reported FHL causative genes encode molecules that are involved in the intracellular trafficking of cytotoxic granules and their fusion to the plasma membrane, followed by the delivery of their contents into the target cell through immunological synapses. However, other components essential for cytotoxic granule degranulation for CTLs are not well understood. To identify indispensable molecules for cytotoxic granule degranulation, we established an efficient in vitro CD8 amplification and degranulation system to perform a CRISPR-screening. To test this system, we transduced OT-I CD8 cells with a gRNA library targeting PIP 3binding proteins, which are important for cell adhesion, subcellular trafficking and T cell activation. Cells were harvested at short (Day 8) and medium-timepoints (Day 15) to evaluate the frequency of gRNAs in the degranulated (CD107+) and non-degranulated fractions. Of the top 10 hits of genes whose deletion suppressed degranulation, five were already listed in the 2022 IUIS classification of inborn error of immunity. We will use individual knockouts to evaluate how these genes affect degranulation. To expand to genome-wide screening, we have also subcloned a full library of sgRNAs into our high efficiency retroviral vector. Thus, we established an efficient assay system to perform a CRISPR-screening for CD8 T cells and can detect genes affected by primary immunodeficiencies. This work was supported by the Intramural Research Program of NIAID, NIH.

Development of colloidal gold immunochromatography and reverse transcription loop-mediated isothermal amplification assays to detect lychnis mottle virus.

Lychnis mottle virus (LycMoV; genus Unassigned, family Secoviridae) infection of Angelica sinensis produces mottle and mosaic symptoms, damaging the host. Early detection of relevant pathogens is the most critical step in preventing the potential transmission of infectious disease. Polyclonal antibodies (pAbs) with high potency and high specificity were prepared using the recombinant LycMoV capsid protein as an antigen. Here, we developed and optimized a rapid colloidal gold immunochromatography assay (GICA) detection system for LycMoV using this antibody. Under optimum conditions, GICA specifically detected (up to 10,000-fold) positive LycMoV samples. A real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) system was also established by selecting the primers with high sensitivity and specificity to LycMoV. The RT-LAMP detection threshold was 1.42 fg/μl (291 copies/μl). A GICA-RT-LAMP assay system was further established and optimized. The minimum GICA detection line was calculated at 1.52x10-2 ng/μl. Although GICA did not detect positive samples after capturing 2.53x10-3 ng/μl of virus, GICA-LAMP and GICA-RT-PCR did, whose sensitivity was comparatively greater than six-fold. This is the first report showing that GICA-RT-LAMP is a cost effective approach for use in detecting LycMoV without extracting nucleic acids. These sensitive assays will help improve virus disease management in A. sinensis crops.

Efficient structural manipulation of PbS in Er-doped silica optical fibers for enhanced amplification systems

In active silica optical fibers, improving the solubility and uniformity of non-concentration-quenching luminescence centers is a key strategy to achieve high gain fiber amplifiers. Herein, we report a novel Er-doped silica optical fiber (EDF) modified by PbS semiconductor incorporation for the first time. Owing to the bridge role of PbS, Er2O3 nanofilms deposited on the substrate via atomic layer deposition (ALD) have porous and loosely layered network structures with an increasing number of ALD-PbS layers. Subsequently, during the high-temperature fiber preparation process, this loose porous structure aids the dissolution and provides an optimal local environment of Er3+ ions in the silica matrix. As a result, the maximum phonon energy of the silica network for PbS/Er co-doped silica optical fiber (PEDF) is reduced, resulting in an increase of the Er3+ ion fluorescence lifetime, approximately 11.455 ms. Furthermore, we achieved 34 dB maximum gain of PEDF sample with the lowest noise figure of 3.9 dB at 1.5 μm. The gain bandwidth over 20 dB of PEDF is approximately 46 nm, which is nearly 44% higher than that of EDF. These results indicate that the PEDF prepared by structural manipulation has great potentials in optical fiber amplifier and laser applications.

Self‐configuration system and frequency characterization of a programmable photodetector ASIC

SummaryIn this work, a self‐configuration system and the frequency characterization for a fully integrated CMOS photodetector sensor is presented. The sensor is composed of pixels with programmable switches that allow each pixel to connect with its neighbors; in this way, an arbitrary detection pattern can be synthesized on it. The design was aimed to be part of an optical encoder based on a non‐diffractive light beam; therefore, the purpose of the self‐configuration routine is to find the center of the incident non‐diffractive beam and then configure the detection pattern around it. The corresponding algorithm is implemented on a Zynq‐7000SoCallowing to automate the alignment of the beam with the detection pattern, without using micrometric positioning procedures. The frequency response of the analog front‐end of the entire chip (the pixels and the amplification system) is addressed via SPICE simulations and experimental data and is consistent with the classical mathematical models, allowing us to propose future improvements to the design.

Chirped pulse amplification based on praseodymium-doped fluoride fibers.

We have developed a chirped pulse amplification system for ultrashort 1300 nm pulses based on praseodymium-doped fluoride fibers (Pr:ZBLAN). The 1300 nm seed pulse is generated through soliton-dispersive wave coupling in a highly nonlinear fiber pumped by a pulse from an erbium-doped fiber laser. The seed pulse is stretched with a grating stretcher to ∼150 ps and amplified with a two-stage Pr:ZBLAN amplifier. The average power reaches ∼112 mW at the repetition rate of 40 MHz. The pulse is compressed to 225 fs by using a pair of gratings without serious phase distortion.