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High-throughput Proteomics Enabled by a Photocleavable Surfactant
摘要： Mass spectrometry (MS)-based proteomics provides unprecedented opportunities for understanding the structure and function of proteins in complex biological systems; however, protein solubility and sample preparation before MS analysis remain a bottleneck preventing high-throughput proteomics. Herein, we report for the first time a high-throughput bottom-up proteomics method enabled by a newly developed MS-compatible photocleavable surfactant, 4-hexylphenylazosulfonate (Azo)[1] that facilitates robust protein extraction, rapid enzymatic digestion (30 min compared to overnight), and subsequent MS-analysis following UV degradation. Moreover, we developed an Azo-aided bottom-up method for efficient analysis of integral membrane proteins, which are key drug targets and are generally underrepresented in global proteomic studies. Furthermore, we demonstrated the unique ability of Azo to serve as an “all-in-one” MS-compatible surfactant for both top-down and bottom-up proteomics, with streamlined workflows for high-throughput proteomics amenable to clinical applications.
关键词： membrane proteomics,high-throughput proteomics,photocleavable surfactant
更新于2025-09-23 15:19:57
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High-throughput label-free molecular fingerprinting flow cytometry
摘要： Flow cytometry is an indispensable tool in biology for counting and analyzing single cells in large heterogeneous populations. However, it predominantly relies on fluorescent labeling to differentiate cells and, hence, comes with several fundamental drawbacks. Here, we present a high-throughput Raman flow cytometer on a microfluidic chip that chemically probes single live cells in a label-free manner. It is based on a rapid-scan Fourier-transform coherent anti-Stokes Raman scattering spectrometer as an optical interrogator, enabling us to obtain the broadband molecular vibrational spectrum of every single cell in the fingerprint region (400 to 1600 cm?1) with a record-high throughput of ~2000 events/s. As a practical application of the method not feasible with conventional flow cytometry, we demonstrate high-throughput label-free single-cell analysis of the astaxanthin productivity and photosynthetic dynamics of Haematococcus lacustris.
关键词： high-throughput,astaxanthin,label-free,single-cell analysis,microfluidics,flow cytometry,Raman spectroscopy,Haematococcus lacustris,coherent anti-Stokes Raman scattering
更新于2025-09-19 17:15:36
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Accelerated Discovery of Two-Dimensional Optoelectronic Octahedral Oxyhalides via High-Throughput <i>Ab Initio</i> Calculations and Machine Learning
摘要： Traditional trial-and-error methods are obstacles for large-scale searching of new optoelectronic materials. Here, we introduce a method combining high-throughput ab initio calculations and machine-learning approaches to predict two-dimensional octahedral oxyhalides with improved optoelectronic properties. We develop an effective machine-learning model based on an expansive data set generated from density functional calculations including the geometric and electronic properties of 300 two-dimensional octahedral oxyhalides. Our model accelerates the screening of potential optoelectronic materials of 5000 two-dimensional octahedral oxyhalides. The distorted stacked octahedral factors proposed in our model play essential roles in the machine-learning prediction. Several potential two-dimensional optoelectronic octahedral oxyhalides with moderate band gaps, high electron mobilities, and ultrahigh absorbance coefficients are successfully hypothesized.
关键词： band gaps,optoelectronic materials,two-dimensional octahedral oxyhalides,absorbance coefficients,electron mobilities,high-throughput ab initio calculations,machine learning
更新于2025-09-19 17:13:59
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Off-colony screening of biosynthetic libraries by rapid laser-enabled mass spectrometry
摘要： Leveraging advances in DNA synthesis and molecular cloning techniques, synthetic biology increasingly makes use of large construct libraries to explore large design spaces. For biosynthetic pathway engineering the ability to screen these libraries for a variety of metabolites of interest is essential. If the metabolite of interest or the metabolic phenotype is not easily measurable, screening soon becomes a major bottleneck involving time-consuming culturing, sample preparation, and extraction. To address this, we demonstrate the use of automated Laser-Assisted Rapid Evaporative Ionisation Mass Spectrometry (LA-REIMS) – a form of ambient laser desorption ionisation mass spectrometry – to perform rapid mass spectrometry analysis direct from agar plate yeast colonies without sample preparation or extraction. We use LA-REIMS to assess production levels of violacein and betulinic acid directly from yeast colonies at a rate of 6 colonies per minute. We then demonstrate the throughput enabled by LA-REIMS by screening over 450 yeast colonies in under 4 hours, while simultaneously generating recoverable glycerol stocks of each colony in real-time. This showcases LA-REIMS as a pre-screening tool to complement downstream quantification methods such as LCMS. Through pre-screening several hundred colonies with LA-REIMS, we successfully isolate and verify a strain with a 2.5-fold improvement in betulinic acid production. Finally, we show that LA-REIMS can detect 20 out of a panel of 27 diverse biological molecules, demonstrating the broad applicability of LA-REIMS to metabolite detection. The rapid and automated nature of LA-REIMS makes this a valuable new technology to complement existing screening technologies currently employed in academic and industrial workflows.
关键词： synthetic biology,metabolic engineering,REIMS,ambient mass spectrometry,high throughput screening
更新于2025-09-19 17:13:59
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Dynamic Multifocus Laser Writing with Acousto‐Optofluidics
摘要： Laser writing of materials is normally performed by the sequential scanning of a single focused beam across a sample. This process is time-consuming and it can severely limit the throughput of laser systems in key applications such as surgery, microelectronics, or manufacturing. A parallelization strategy based on ultrasound waves in a liquid to diffract light into multiple beamlets is reported. Adjusting amplitude, frequency, or phase of ultrasound allows tunable multifocus distributions with sub-microsecond control. When combined with sample translation, the dynamic splitting of light leads to high-throughput laser processing, as demonstrated by locally modifying the morphological and wettability properties of metals, polymers, and ceramics. The results illustrate how acousto-optofluidic systems are universal tools for fast multifocus generation, with potential impact in fields such as imaging or optical trapping.
关键词： complex patterning,laser ablation,high throughput processing,wettability
更新于2025-09-19 17:13:59
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Vacuum-Deposited Blue Inorganic Perovskite Light-Emitting Diodes
摘要： Perovskite light-emitting diodes (PeLEDs) have drawn great research attention due to their outstanding electroluminescence performance by solution processing. PeLEDs made by thermal evaporation are relatively rarely explored but compatible to existing OLED industrial lines. Blue-emitting PeLEDs are all based on organic-containing perovskites, rather than more stable all-inorganic perovskites because of their poor solubility, too fast crystallization, uneven discrete film and unattainable pure blue emission. Here we report all-inorganic, vacuum processed blue PeLEDs. High-throughput combinatorial approaches are employed to optimize Cs:Pb:Br:Cl composition in our dual-source co-evaporation system to achieve the balance between film photoluminescence and injection efficiency. As-deposited perovskite films demonstrated excellent intrinsic stability against heat, UV-light and humidity attack. A series of PeLEDs were obtained covering standard blue spectral region with a best luminance of 121 cd/m2 as well as an EQE of 0.38%. We believe the vacuum processing strategy demonstrated here provide a very promising alternative way to produce efficient and stable all-inorganic blue-emitting PeLEDs.
关键词： all-inorganic perovskite,stability,high-throughput combinatorial approaches,vacuum-deposited,blue-emitting
更新于2025-09-16 10:30:52
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Probing Specificity of Protein–Protein Interactions with Chiral Plasmonic Nanostructures
摘要： Protein?protein interactions (PPIs) play a pivotal role in many biological processes. Discriminating functionally important well-defined protein?protein complexes formed by specific interactions from random aggregates produced by nonspecific interactions is therefore a critical capability. While there are many techniques which enable rapid screening of binding affinities in PPIs, there is no generic spectroscopic phenomenon which provides rapid characterization of the structure of protein?protein complexes. In this study we show that chiral plasmonic fields probe the structural order and hence the level of PPI specificity in a model antibody?antigen system. Using surface-immobilized Fab′ fragments of polyclonal rabbit IgG antibodies with high specificity for bovine serum albumin (BSA), we show that chiral plasmonic fields can discriminate between a structurally anisotropic ensemble of BSA-Fab′ complexes and random ovalbumin (OVA)-Fab′ aggregates, demonstrating their potential as the basis of a useful proteomic technology for the initial rapid high-throughput screening of PPIs.
关键词： specificity,Protein?protein interactions,structural order,chiral plasmonic nanostructures,high-throughput screening
更新于2025-09-16 10:30:52
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High-throughput analysis of single particles by micro laser induced breakdown spectroscopy
摘要： Multi-elemental analysis of particulate matters is highly desirable for the identification of their emission sources. However, the traditional techniques for single particle analysis usually require time-consuming sample preparation procedures. We report here a system that combines bright-field microscopy with laser induced breakdown spectroscopy (LIBS), allowing rapid and multi-elemental analysis of single particles. The improved throughput of the system was benefited from the use of ultra-thin polyethylene films for particle immobilization such that their spectra were collected successively. The simple composition of the films avoided their interferences in the particles’ LIBS spectra. Moreover, the small thickness of the films allowed their rapid vaporization during laser ablation such that the plasma emissions from the particles were not quenched by their substrates. The performances of the system were demonstrated by studying the heterogeneity in particles’ compositions and discriminating dust particles with very similar chemicals, suggesting that it is a promising tool for environmental risk assessment.
关键词： single particles,Laser induced breakdown spectroscopy,high-throughput
更新于2025-09-16 10:30:52
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Rapid Assessment of Water Toxicity by Plasmonic Nanomechanical Sensing
摘要： The ability to rapidly and accurately detect water toxicity is crucial for monitoring water quality and assessing toxic risk, but such detection remains a great challenge. Here, we present a plasmonic nanomechanical sensing (PNMS) system for the rapid assessment of water toxicity. This technique is based on the plasmonic sensing of the nanomechanical movement of single bacterial cells, which could be inhibited upon exposure to potential toxicants. By correlating the amplitude of nanomechanical movement with bacterial activity, we detected a variety of toxic substances in water. The direct readout of bacterial activity via PNMS allowed for a high sensitivity to toxicants in water, thereby enabling us to evaluate the acute toxicological effect of chemical compounds rapidly. The PNMS method is promising for online alerts of water quality safety and for assessing chemical hazards. We anticipate that PNMS is also suitable for a wide range of other applications, including bacterial detection and high-throughput screening of anti-bacterial materials.
关键词： water toxicity,nanomechanical movement,high-throughput,detection,Single cell
更新于2025-09-12 10:27:22
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Efficient Exploration of the Composition Space in Ternary Organic Solar Cells by Combining High‐Throughput Material Libraries and Hyperspectral Imaging
摘要： Organic solar cells based on ternary active layers can lead to higher power conversion efficiencies than corresponding binaries, and improved stability. The parameter space for optimization of multicomponent systems is considerably more complex than that of binaries, due to both, a larger number of parameters (e.g., two relative compositions rather than one) and intricate morphology–property correlations. Most experimental reports to date reasonably limit themselves to a relatively narrow subset of compositions (e.g., the 1:1 donor/s:acceptor/s trajectory). This work advances a methodology that allows exploration of a large fraction of the ternary phase space employing only a few (<10) samples. Each sample is produced by a designed sequential deposition of the constituent inks, and results in compositions gradients with ≈5000 points/sample that cover about 15%–25% of the phase space. These effective ternary libraries are then colocally imaged by a combination of photovoltaic techniques (laser and white light photocurrent maps) and spectroscopic techniques (Raman, photoluminescence, absorption). The generality of the methodology is demonstrated by investigating three ternary systems, namely PBDB-T:ITIC:PC70BM, PTB7-Th:ITIC:PC70BM, and P3HT:O-IDFBR:O-IDTBR. Complex performance-structure landscapes through the ternary diagram as well as the emergence of several performance maxima are discovered.
关键词： high-throughput screening,layer-by-layer deposition,organic solar cells,ternaries,Raman imaging
更新于2025-09-12 10:27:22