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At least 307 records · Page 17

Multiplex base editing to convert $\mathrm{TAG}$ into $\mathrm{TAA}$ codons in the human genome

Whole-genome recoding has been shown to enable nonstandard amino acids, biocontainment and viral resistance in bacteria. Here we take the first steps to extend this to human cells demonstrating exceptional base editing to convert TAG to TAA for 33 essential genes via a single transfection, and examine base-editing genome-wide (observing ~40 C-to-T off-target events in essential gene exons). We also introduce GRIT, a computational tool for recoding. This demonstrates the feasibility of recoding, and highly multiplex editing in mammalian cells.

59 BASIC BIOLOGICAL SCIENCES↗

Substrate multiplexed protein engineering facilitates promiscuous biocatalytic synthesis

Enzymes with high activity are readily produced through protein engineering, but intentionally and efficiently engineering enzymes for an expanded substrate scope is a contemporary challenge. One approach to address this challenge is Substrate Multiplexed Screening (SUMS), where enzyme activity is measured on competing substrates. SUMS has long been used to rigorously quantitate native enzyme specificity, primarily for in vivo settings. SUMS has more recently found sporadic use as a protein engineering approach but has not been widely adopted by the field, despite its potential utility. Here, we develop principles of how to design and interpret SUMS assays to guide protein engineering. This rich information enables improving activity with multiple substrates simultaneously, identifies enzyme variants with altered scope, and indicates potential mutational hot-spots as sites for further engineering. These advances leverage common laboratory equipment and represent a highly accessible and customizable method for enzyme engineering.

59 BASIC BIOLOGICAL SCIENCES↗

A substrate-multiplexed platform for profiling enzymatic potential of plant family 1 glycosyltransferases

Plants have expanded various biosynthetic enzyme families to produce a wide diversity of natural products; however, most enzymes encoded in plant genomes remain uncharacterized, highlighting the need for new functional genomic approaches. Here, we report a platform enabling the rapid functional characterization of plant family 1 glycosyltransferases, which serve important roles in plant development, defense, and communication. Using substrate-multiplexed reactions, mass spectrometry, and automated analysis, we screen 85 enzymes against a diverse library of 453 natural products, for a total of nearly 40,000 possible reactions. The resulting dataset reveals a widespread promiscuity and a strong preference for planar, hydroxylated aromatic substrates among family 1 glycosyltransferases. We also characterize glycosyltransferases with an unusually wide substrate scope and with a non-canonical Cys-Asp catalytic dyad. This work establishes a widely-applicable enzymatic screening pipeline, reflects the immense glycosylation capability of plants, and has implications in biocatalysis, metabolic engineering, and gene discovery.

Sirirungruang, Sasilada↗

Reproducible workflow for multiplexed deep-scale proteome and phosphoproteome analysis of tumor tissues by liquid chromatography–mass spectrometry

Here we present an optimized workflow for global proteome and phosphoproteome analysis of tissues or cell lines that uses isobaric tags (TMT (tandem mass tags)-10) for multiplexed analysis and relative quantification, and provides 3×higher throughput than iTRAQ (isobaric tags for absolute and relative quantification)-4-based methods with high intraand inter-laboratory reproducibility. The workflow was systematically characterized and benchmarked across three independent laboratories using two distinct breast cancer subtypes from patient-derived xenograft models to enable assessment of proteome and phosphoproteome depth and quantitative reproducibility. Each plex consisted of ten samples, each being 300 µg of peptide derived from <50 mg of wet-weight tissue. Of the 10,000 proteins quantified per sample, we could distinguish 7,700 human proteins derived from tumor cells and 3100 mouse proteins derived from the surrounding stroma and blood. The maximum deviation across replicates and laboratories was <7%, and the interlaboratory correlation for TMT ratio–based comparison of the two breast cancer subtypes was r > 0.88. The maximum deviation for the phosphoproteome coverage was <24% across laboratories, with an average of >37,000 quantified phosphosites per sample and differential quantification correlations of r > 0.72. The full procedure, including sample processing and data generation, can be completed within 10 d for ten tissue samples, and 100 samples can be analyzed in ~4 months using a single LC-MS/MS instrument. The high quality, depth, and reproducibility of the data obtained both within and across laboratories should enable new biological insights to be obtained from mass spectrometry-based proteomics analyses of cells and tissues together with proteogenomic data integration.

Mertins, Philipp↗

Multiwavelength metasurfaces through spatial multiplexing

Metasurfaces are two-dimensional arrangements of optical scatterers rationally arranged to control optical wavefronts. Despite the significant advances made in wavefront engineering through metasurfaces, most of these devices are designed for and operate at a single wavelength. Here we show that spatial multiplexing schemes can be applied to increase the number of operation wavelengths. We use a high contrast dielectric transmittarray platform with amorphous silicon nano-posts to demonstrate polarization insensitive metasurface lenses with a numerical aperture of 0.46, that focus light at 915 and 1550 nm to the same focal distance. We investigate two different methods, one based on large scale segmentation and one on meta-atom interleaving, and compare their performances. An important feature of this method is its simple generalization to adding more wavelengths or new functionalities to a device. Furthermore, it provides a relatively straightforward method for achieving multi-functional and multiwavelength metasurface devices.

36 MATERIALS SCIENCE↗

Orbital Angular Momentum-based Space Division Multiplexing for High-capacity Underwater Optical Communications

To increase system capacity of underwater optical communications, we employ the spatial domain to simultaneously transmit multiple orthogonal spatial beams, each carrying an independent data channel. In this paper, we show up to a 40-Gbit/s link by multiplexing and transmitting four green orbital angular momentum (OAM) beams through a single aperture. Moreover, we investigate the degrading effects of scattering/turbidity, water current, and thermal gradient-induced turbulence, and we find that thermal gradients cause the most distortions and turbidity causes the most loss. We show systems results using two different data generation techniques, one at 1064 nm for 10-Gbit/s/beam and one at 520 nm for 1-Gbit/s/beam; we use both techniques since present data-modulation technologies are faster for infrared (IR) than for green. For the 40-Gbit/s link, data is modulated in the IR, and OAM imprinting is performed in the green using a specially-designed metasurface phase mask. For the 4-Gbit/s link, a green laser diode is directly modulated. Lastly, we show that inter-channel crosstalk induced by thermal gradients can be mitigated using multi-channel equalisation processing.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

High-resolution gamma-ray spectroscopy with a microwave-multiplexed transition-edge sensor array

We demonstrate very high resolution photon spectroscopy with a microwave-multiplexed two-pixel transition-edge sensor (TES) array. We measured a {sup 153}Gd photon source and achieved an energy resolution of 63 eV full-width-at-half-maximum at 97 keV and an equivalent readout system noise of 86 pA/√(Hz) at the TES. The readout circuit consists of superconducting microwave resonators coupled to radio-frequency superconducting-quantum-interference-devices and transduces changes in input current to changes in phase of a microwave signal. We use flux-ramp modulation to linearize the response and evade low-frequency noise. This demonstration establishes one path for the readout of cryogenic X-ray and gamma-ray sensor arrays with more than 10{sup 3} elements and spectral resolving powers R=λ/Δλ>10{sup 3}.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

Subwavelength light focusing with a single slit lens based on the spatial multiplexing of chirped surface gratings

A lens consisting of a subwavelength slit engraved into a metal film and surrounded by periodic surface gratings and spatial multiplexing chirped surface gratings (SMCSGs) for focusing light is proposed. The focal length of the lens can be accurately designed and tuned by controlling the periods of the local gratings of the SMCSGs. Simulation results show that a subwavelength beam spot can be produced by the lens at a distance of several times the incident wavelength from the slit and the difference between the simulated and designed focal lengths can be reduced to below 3% of the designed focal length.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

Multiplexed absorption tomography with calibration-free wavelength modulation spectroscopy

We propose a multiplexed absorption tomography technique, which uses calibration-free wavelength modulation spectroscopy with tunable semiconductor lasers for the simultaneous imaging of temperature and species concentration in harsh combustion environments. Compared with the commonly used direct absorption spectroscopy (DAS) counterpart, the present variant enjoys better signal-to-noise ratios and requires no baseline fitting, a particularly desirable feature for high-pressure applications, where adjacent absorption features overlap and interfere severely. We present proof-of-concept numerical demonstrations of the technique using realistic phantom models of harsh combustion environments and prove that the proposed techniques outperform currently available tomography techniques based on DAS.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

A near-infrared 64-pixel superconducting nanowire single photon detector array with integrated multiplexed readout

We demonstrate a 64-pixel free-space-coupled array of superconducting nanowire single photon detectors optimized for high detection efficiency in the near-infrared range. An integrated, readily scalable, multiplexed readout scheme is employed to reduce the number of readout lines to 16. The cryogenic, optical, and electronic packaging to read out the array as well as characterization measurements are discussed.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

Accelerated nanoscale magnetic resonance imaging through phase multiplexing

We report a method for accelerated nanoscale nuclear magnetic resonance imaging by detecting several signals in parallel. Our technique relies on phase multiplexing, where the signals from different nuclear spin ensembles are encoded in the phase of an ultrasensitive magnetic detector. We demonstrate this technique by simultaneously acquiring statistically polarized spin signals from two different nuclear species ({sup 1}H, {sup 19}F) and from up to six spatial locations in a nanowire test sample using a magnetic resonance force microscope. We obtain one-dimensional imaging resolution better than 5 nm, and subnanometer positional accuracy.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

Multiplexed infrared photodetection using resonant radio-frequency circuits

We demonstrate a room-temperature semiconductor-based photodetector where readout is achieved using a resonant radio-frequency (RF) circuit consisting of a microstrip split-ring resonator coupled to a microstrip busline, fabricated on a semiconductor substrate. The RF resonant circuits are characterized at RF frequencies as function of resonator geometry, as well as for their response to incident IR radiation. The detectors are modeled analytically and using commercial simulation software, with good agreement to our experimental results. Though the detector sensitivity is weak, the detector architecture offers the potential for multiplexing arrays of detectors on a single read-out line, in addition to high speed response for either direct coupling of optical signals to RF circuitry, or alternatively, carrier dynamics characterization of semiconductor, or other, material systems.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

Pixel multiplexing technique for real-time three-dimensional-imaging laser detection and ranging system using four linear-mode avalanche photodiodes

The avalanche-photodiode-array (APD-array) laser detection and ranging (LADAR) system has been continually developed owing to its superiority of nonscanning, large field of view, high sensitivity, and high precision. However, how to achieve higher-efficient detection and better integration of the LADAR system for real-time three-dimensional (3D) imaging continues to be a problem. In this study, a novel LADAR system using four linear mode APDs (LmAPDs) is developed for high-efficient detection by adopting a modulation and multiplexing technique. Furthermore, an automatic control system for the array LADAR system is proposed and designed by applying the virtual instrumentation technique. The control system aims to achieve four functions: synchronization of laser emission and rotating platform, multi-channel synchronous data acquisition, real-time Ethernet upper monitoring, and real-time signal processing and 3D visualization. The structure and principle of the complete system are described in the paper. The experimental results demonstrate that the LADAR system is capable of achieving real-time 3D imaging on an omnidirectional rotating platform under the control of the virtual instrumentation system. The automatic imaging LADAR system utilized only 4 LmAPDs to achieve 256-pixel-per-frame detection with by employing 64-bit demodulator. Moreover, the lateral resolution is ∼15 cm and range accuracy is ∼4 cm root-mean-square error at a distance of ∼40 m.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

Multiplexed high resolution soft x-ray RIXS

High-resolution Resonance Inelastic X-ray Scattering (RIXS) is a technique that allows us to probe the electronic excitations of complex materials with unprecedented precision. However, the RIXS process has a low cross section, compounded by the fact that the optical spectrometers used to analyze the scattered photons can only collect a small solid angle and overall have a small efficiency. Here we present a method to significantly increase the throughput of RIXS systems, by energy multiplexing, so that a complete RIXS map of scattered intensity versus photon energy in and photon energy out can be recorded simultaneously{sup 1}. This parallel acquisition scheme should provide a gain in throughput of over 100.. A system based on this principle, QERLIN, is under construction at the Advanced Light Source (ALS).

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

Spin-Wave Frequency Division Multiplexing in an Yttrium Iron Garnet Microstripe Magnetized by Inhomogeneous Field.

Spin waves are promising candidates for information processing and transmission in a broad frequency range. In the realization of magnonic devices, the frequency related division of the spin waves is a critical function for parallel information processing. In this work, we demonstrate a proof-of-concept spin-wave frequency division multiplexing method by magnetizing a homogeneous magnetic microstripe with an inhomogeneous field. The symmetry breaking additional field is introduced by a Permalloy stripe simply placed in lateral proximity to an yttrium iron garnet waveguide. Spin waves with different frequencies can propagate independently, simultaneously, and separately in space along the shared waveguide. This work demonstrates one potential way for parallel information transmission and processing in magnonics.

Zhang, Zhizhi↗

Simulations of wavelength-multiplexed holography for single-shot spatiotemporal characterization of NIF’s advanced radiographic capability (ARC) laser

We simulate the use of a newly developed single-shot wavelength-multiplexed holography-based diagnostic, STRIPED FISH, to fully characterize the as-delivered laser pulses of the National Ignition Facility’s Advanced Radiographic Capability (NIF-ARC) laser. To that end, we have performed simulations of the NIF-ARC pulse incorporating (a) a time-integrated spatial-profile measurement and a complete temporal-intensity-and-phase measurement using a frequency resolved optical gating, but without any spatiotemporal pulse characterizations, and (b) simulated first-order spatiotemporal distortions, which could be measured on a single shot if a STRIPED FISH device were deployed.

47 OTHER INSTRUMENTATION↗

Semiconductor laser with a birefringent external cavity for information systems with wavelength division multiplexing

The spectrum of a semiconductor laser with a birefringent external Gires – Tournois cavity is studied. The generation of two main laser modes corresponding to the ordinary and extraordinary wave resonances is found. It is shown that the radiation spectrum is controlled with a high energy efficiency without losses for spectral filtration. The possibility of using two-mode lasing in optical communication systems with wavelength division multiplexing is shown. (control of laser radiation parameters)

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗