High-throughput Testing Research Articles

Revealing the Phenotypic and Genomic Background for PHA Production from Rapeseed-Biodiesel Crude Glycerol Using Photobacterium ganghwense C2.2.

Polyhydroxyalkanoates (PHA) are promising biodegradable and biocompatible bioplastics, and extensive knowledge of the employed bacterial strain's metabolic capabilities is necessary in choosing economically feasible production conditions. This study aimed to create an in-depth view of the utilization of Photobacterium ganghwense C2.2 for PHA production by linking a wide array of characterization methods: metabolic pathway annotation from the strain's complete genome, high-throughput phenotypic tests, and biomass analyses through plate-based assays and flask and bioreactor cultivations. We confirmed, in PHA production conditions, urea catabolization, fatty acid degradation and synthesis, and high pH variation and osmotic stress tolerance. With urea as a nitrogen source, pure and rapeseed-biodiesel crude glycerol were analyzed comparatively as carbon sources for fermentation at 20 °C. Flask cultivations yielded 2.2 g/L and 2 g/L PHA at 120 h, respectively, with molecular weights of 428,629 g/mol and 81,515 g/mol. Bioreactor batch cultivation doubled biomass accumulation (10 g/L and 13.2 g/L) in 48 h, with a PHA productivity of 0.133 g/(L·h) and 0.05 g/(L·h). Thus, phenotypic and genomic analyses determined the successful use of Photobacterium ganghwense C2.2 for PHA production using urea and crude glycerol and 20 g/L NaCl, without pH adjustment, providing the basis for a viable fermentation process.

Application of Real-Time Cell Analysis Biosensor Technology for Drug Cytotoxicity Studies in Primary Lung Cancer Cells

Biosensing technologies can be used for toxicity studies on cell cultures. The xCELLigence Real-Time Cell Analysis (RTCA) system is a portable label-free detection method with the capability to analyze cellular proliferation, migration, invasion, growth, morphology states, and cytotoxicity of adherent cells in a non-invasive environment. In pharmacological context, most xCELLigence studies were done using cancer cell lines. Hypothetically, implementing the system with primary cell culture can anticipate actions in vivo better than those seen on cell lines. This could potentially provide useful information on how the drug responds to targeted cancer. In this work, we explore the usage of biosensing technology to establish a concept for high-throughput testing of chemotherapeutics agents on primary patient-derived cancer cells. Four self-established primary lung cancer cells (samples A36, A38, A39, and A40) were derived from biopsies of patients, cultured, and tested with cisplatin. The xCELLigence was used to monitor the response of primary lung cancer cells toward different concentrations of cisplatin ranging from 1 to 120 μM. Cisplatin was found to be effective at approximately 20 μM after 48 h of exposure with A38 and A39 showing drug resistance behavior at below 20 μM concentration. The IC50 values were between 22.88 to 76.22 μM. This indicates that the RTCA biosensor technology has good potential in predicting the cell’s response to chemotherapy especially when primary cancer cells are used. This approach can be the fundamental reference in the future to support personalized chemotherapy treatment by providing additional information on metastatic behavior, cytotoxic and drug-resistance effects of the cancer cells.Graphical Abstract

Recent advances in the use of graphene for sample preparation

Sample preparation is playing an increasingly important role in sample analysis. The enrichment efficiency of the target and the removal effect of the sample matrix are strongly dependent on the extraction material. Therefore, the development of efficient extraction materials is an important research focus in the field of sample preparation. Various advanced materials such as nanomaterials, mesoporous materials, ionic liquids, aerogels, carbon materials, metal-organic frameworks, and covalent organic frameworks have been introduced to produce a diverse range of extraction materials for sample preparation. Owing to its unique physical and chemical properties, graphene, an excellent carbon nanomaterial, has attracted significant attention in different areas. Due to their unique advantages of large surface area, large π-electrons, excellent adsorption properties, abundant functional groups, and facile chemical modification, graphene-based materials have displayed excellent extraction performance for diverse analytes. Furthermore, graphene-based extraction materials have been applied to pretreat real samples from different fields. This paper provides an overview of the recent advances in graphene sample preparation from 2020 to date. The manuscript covers the use of graphene, graphene oxide, and the related functionalized materials as sorbents, as well as their specific applications in cartridge solid-phase extraction, dispersive solid-phase extraction, magnetic solid-phase extraction, stir bar sorptive extraction, fiber solid-phase microextraction, and in-tube solid-phase microextraction. To prevent the aggregation of graphene, three-dimensional graphene, porous graphene aerogels, graphene-modified silica, and stainless-steel mesh were developed for cartridge solid-phase extraction. Furthermore, some graphene-based extraction materials were used to develop online solid-phase extraction, which allowed for automatic and high-throughput tests. Graphene nanosheets and their hybrid materials with molybdenum disulfide or zinc oxide nanoparticles have been applied to dispersive solid-phase extraction, and several types of contaminants, including metal ions, bisphenol endocrine disruptors, paraben preservatives, and phthalates, could be captured. By combination with magnetic materials using the coprecipitation method or via chemical post-modification, many magnetic graphene extraction materials have been produced for magnetic solid-phase extraction. The introduction of magnetic graphene not only enhanced the extraction efficiency but also simplified the test process, making it highly suitable for complex samples such as food and biological samples. Similar to magnetic solid-phase extraction, stir bar sorptive extraction is a very simple and efficient extraction method that shows good extraction performance for metal ions and organic pollutants from environmental water, medicines in urine, and organic pollutants in cosmetics. In addition to its excellent applicability to solid-phase extraction, graphene delivered satisfactory performance for solid-phase microextraction. Graphene has been used as an extraction coating for the extraction of fibers or tubes by coupling solid-phase microextraction with chromatographic detection, and many kinds of organic pollutants, including polychlorinated biphenyls, phthalates, polycyclic aromatic hydrocarbons, toluene, xylenes, organophosphorus pesticides, phenoxy acid herbicides, and antibiotics, in environmental or biological samples have been successfully determined. The extraction mechanism, including π-π, electrostatic, hydrophobic, hydrophilic, and hydrogen-bonding interactions, is also discussed. Because of the mixed-mode interactions and rich functionalization, graphene-based extraction materials could effectively capture and selectively enrich different types of species. These extraction or microextraction techniques have been coupled with detection methods such as chromatography, mass spectrometry, and atomic absorption spectroscopy and widely used in environmental monitoring, food safety, and biochemical analysis. The future development of graphene in the field of sample pretreatment focuses on the following aspects: 1) functionalization of graphene with specific groups such as affinity groups, chelating groups, and molecularly imprinted sites to achieve unique extraction selectivity; 2) combination of graphene with the advanced materials, including covalent organic frameworks, metal organic frameworks, aerogels, and nanomaterials, thus realizing the complementary advantages between materials, so that the hybrid graphene materials find broad application prospects in sample preparation; 3) combination of electromagnetic materials with graphene to form electromagnetic composites, as well as the use of electromagnetic fields to improve extraction selectivity and efficiency; 4) exploiting the good performance of graphene-based materials to overcome the difficulty encountered in the pretreatment of complex samples; 5) development of more green methods to prepare graphene-based extraction materials or functionalize graphene, in line with the trends in green chemistry; 6) application of more graphene-based materials to online sample preparation for meeting the development trends in the field of analytical chemistry.

Functionalized Titanium Dioxide Nanoparticle-Based Electrochemical Immunosensor for Detection of SARS-CoV-2 Antibody.

The advancement in biosensors can overcome the challenges faced by conventional diagnostic techniques for the detection of the highly infectious severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Hence, the development of an accurate, rapid, sensitive, and selective diagnostic technique can mitigate adverse health conditions caused by SARS-CoV-2. This work proposes the development of an electrochemical immunosensor based on bio-nanocomposites for the sensitive detection of SARS-CoV-2 antibodies through the differential pulse voltammetry (DPV) electroanalytical method. The facile synthesis of chitosan-functionalized titanium dioxide nanoparticles (TiO2-CS bio-nanocomposites) is performed using the sol-gel method. Characterization of the TiO2-CS bio-nanocomposite is accomplished using UV-vis spectroscopy, Raman spectroscopy, X-ray diffraction (XRD), and transmission electron microscopy (TEM). The electrochemical performance is studied using cyclic voltammetry (CV), DPV, and electrochemical impedance spectroscopy (EIS) for its electroanalytical and biosensing capabilities. The developed immunosensing platform has a high sensitivity with a wide range of detection from 50 ag mL-1 to 1 ng mL-1. The detection limit of the SARS-CoV-2 antibody in buffer media is obtained to be 3.42 ag mL-1 and the limit of quantitation (LOQ) to be 10.38 ag mL-1. The electrochemical immunosensor has high selectivity in different interfering analytes and is stable for 10 days. The results suggest that the developed electrochemical immunosensor can be applicable for real sample analysis and further high-throughput testing.

An in vitro alveolar model allows for the rapid assessment of chemical respiratory sensitization with modifiable biomarker endpoints

Diisocyanates are commonly used in polyurethanes where use includes industrial, commercial, and residential applications and can exist as respirable contaminants. These respirable contaminants exist in the air we breathe. Yet, there is no rapid assay available to test for potential respiratory sensitizers. To assess these hazards, as well as to decrease animal numbers used in testing, investigations that lead to verifiable in vitro methods are needed. We describe an easy, reliable, verified cell culture model that can be adopted by any lab capable of performing molecular toxicology. The architecturally relevant alveolar model consists of epithelial cells, macrophage cells, and dendritic cells in a simply maintained submerged system ideal for high-throughput testing. Exposures to contaminants that verify biomarker identification include a known pulmonary sensitizer (isophorone diisocyanate) and a positive control for cellular activation (phorbol 12-myristate 13-acetate/ionomycin). The mitochondrial reactive oxygen species generation and cytostructural changes were assessed with confocal laser scanning microscopy; cell morphology was assessed with scanning electron microscopy; biochemical reactions were assessed via protein arrays; genetic alterations were assessed via gene arrays; and cell surface activation markers were assessed via flow cytometry. Results showed that compared to untreated cultures, isophorone diisocyanate increased markers for dendritic cell activation, trafficking, and antigen presentation; number and length of dendritic protrusions; oxidative stress; and genetic and cytokine expression of neutrophil chemoattractants. The chemokines and cytokines CCL7, CXCL5, IL-6, and IL-8 were identified as biomarkers indicative of respiratory sensitization. By including multiple methods to assess endpoints, the in vitro model described can serve as a high-throughput assay to identify substances which may lead to respiratory sensitization.

Use of compressed sensing to expedite high-throughput diagnostic testing for COVID-19 and beyond.

The rapid spread of SARS-CoV-2 has placed a significant burden on public health systems to provide swift and accurate diagnostic testing highlighting the critical need for innovative testing approaches for future pandemics. In this study, we present a novel sample pooling procedure based on compressed sensing theory to accurately identify virally infected patients at high prevalence rates utilizing an innovative viral RNA extraction process to minimize sample dilution. At prevalence rates ranging from 0-14.3%, the number of tests required to identify the infection status of all patients was reduced by 69.26% as compared to conventional testing in primary human SARS-CoV-2 nasopharyngeal swabs and a coronavirus model system. Our method provided quantification of individual sample viral load within a pool as well as a binary positive-negative result. Additionally, our modified pooling and RNA extraction process minimized sample dilution which remained constant as pool sizes increased. Compressed sensing can be adapted to a wide variety of diagnostic testing applications to increase throughput for routine laboratory testing as well as a means to increase testing capacity to combat future pandemics.

Occurrence, sources, and risk assessment of pyrethroid insecticides in surface water and tap water from Taihu Lake, China

Pyrethroid insecticides are one of the most widely used insecticides globally, posing a severe threat to human health and the environment. In this study, we applied high-throughput organic analysis testing combined with high-volume solid-phase extraction (Hi-throat/Hi-volume SPE) to elucidate the occurrence of 11 pyrethroid insecticides in lake water (n = 37), tributary river water (n = 15), and tap water (n = 6) in the Taihu Lake Basin. Permethrin was found to be the major contributing pyrethroid insecticide (detection rate = 100%). The concentrations of pyrethroid insecticides from different lake regions were revealed in the following descending order: southern > eastern > western > northern. The principal component analysis and multiple linear regression demonstrated that landscape maintenance, agricultural cultivation, and livestock breeding were the main sources of pyrethroid insecticides in the Taihu Lake surface water. Moreover, runoff input plays an important role in their accumulation, while the surrounding rivers contribute 2292 kg of pyrethroid insecticides to Taihu Lake annually. The risk assessment analysis demonstrated that pyrethroid insecticides pose a high risk to both the ecological environment and the surrounding human populations, thereby necessitating effective countermeasures. Furthermore, the pyrethroid insecticides in the Yangtze River Delta region have to be controlled. Overall, this is the first study focused on China that revealed the residue levels in water sources and tap water.

Chondrogenesis of Adipose-Derived Stem Cells Using an Arrayed Spheroid Format.

The chondrogenic response of adipose-derived stem cells (ASCs) is often assessed using 3D micromass protocols that use upwards of hundreds of thousands of cells. Scaling these systems up for high-throughput testing is technically challenging and wasteful given the necessary cell numbers and reagent volumes. However, adopting microscale spheroid cultures for this purpose shows promise. Spheroid systems work with only thousands of cells and microliters of medium. Molded agarose microwells were fabricated using 2% w/v molten agarose and then equilibrated in medium prior to introducing cells. ASCs were seeded at 50, 500, 5k cells/microwell; 5k, 50k, cells/well plate; and 50k and 250k cells/15mL centrifuge tube to compare chondrogenic responses across spheroid and micromass sizes. Cells were cultured in control or chondrogenic induction media. ASCs coalesced into spheroids/pellets and were cultured at 37°C and 5% CO2 for 21days with media changes every other day. All culture conditions supported growth of ASCs and formation of viable cell spheroids/micromasses. More robust growth was observed in chondrogenic conditions. Sulfated glycosaminoglycans and collagen II, molecules characteristics of chondrogenesis, were prevalent in both 5000-cell spheroids and 250,000-cell micromasses. Deposition of collagen I, characteristic of fibrocartilage, was more prevalent in the large micromasses than small spheroids. Chondrogenic differentiation was consistently induced using high-throughput spheroid formats, particularly when seeding at cell densities of 5000 cells/spheroid. This opens possibilities for highly arrayed experiments investigating tissue repair and remodeling during or after exposure to drugs, toxins, or other chemicals. The online version contains supplementary material available at 10.1007/s12195-022-00746-8.

Omics-based high throughput toxicity screening and risk assessment of pollutants

<p indent="0mm">Screening priority pollutants with high toxicity and risk potential from a large number of environmental chemical pollutants is the prerequisite for prevention and control of environmental risks. After entering the living system, toxic pollutants can induce adverse outcomes through a series of disturbed molecular targets and biological pathways. Existing high-throughput toxicity screening methods rely on few and isolated cellular or molecular endpoints, making it difficult to capture the key biological processes that cause toxicity and to determine the direct relationship between chemical pollutants and disease. Meanwhile, the awareness is increasing that we are exposed to a true cocktail of chemicals, among which only a fraction has been identified through analytical methods. Integrated bioanalytical approaches, such as effect-directed analysis and toxicity identification evaluation (TIE), have been used to identify risk drivers in complex mixture pollutants. However, it is still difficult to elucidate the molecular toxicological drivers of chemical stress in the environment. Toxicogenomics take advantage of genome-wide coverage, high throughput, and big data capacity that are associated with omics technologies to achieve technology innovation in screening and risk assessment of chemical pollutants. We reviewed the technical advantages and the recent progress of toxicogenomic methodology in high-throughput toxicological screening and risk assessment of chemical pollutants from four aspects with specific research cases. To identify the molecular events initiated by toxic pollutants, toxicogenomics could uncover a large number of molecular events and pathways potentially disturbed at the early development stage of toxicity. Dose-dependent transcriptomes for high-throughput chemical testing have been developed to quantitatively assess responses induced by chemical pollutants at the molecular level, which improve the sensitivity of identifying bioactivity by 1–3 orders of magnitude. To uncover the biological pathways of adverse health outcome by chemical pollutants, systemic toxicological assessment by omics strategies provide mechanistic information for chemical pollution screening and health risk assessment at multiple biological levels. Through biological pathway enrichment analysis, such as the gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) pathway, the molecular mechanistic information of toxicity could be captured. Moreover, multi-omics integration allows for multi-scale, comprehensive biological network analysis, which provide a systematic understanding of toxicity pathways or adverse outcome pathways (AOPs) to guide ecological and human health risk assessments. For human diseases diagnostics, functional genomics provide a new tool to explore mechanisms of individual disease risk and susceptibility under toxic pollutants exposure. Variations of individual susceptibility to toxicant-induced disease are largely due to genetic susceptibility across different populations. The direct relationship among pollutants, genes and adverse outcomes could be established by combining functional genomics and molecular epidemiological approaches. Finally, for risk management of complex mixture pollutants, it is important to assess the overall toxicity effects of the pool of pollutants and identify the key toxic pollutant. The combination of high-throughput bioassays and chemical analysis can be used to identify the highly bioactive fraction and to screen the key chemical pollutant that pose ecological and health risks. Recently, progresses have been made in integrating transcriptomic techniques with molecular toxicity identification and evaluation (mTIE) to assess the overall effects of mixtures and distinguish the key pollutants in inducing the corresponding biological responses. Particularly, dose-response transcriptome could be used to quantitatively evaluate the overall toxic effects and to guide the key toxicant identification by high mechanistic resolution. In summary, the development of omics-based high-throughput toxicity screening technologies will strongly support accurate environmental risk assessment of conventional and new pollutants in the future.

High-Throughput Dispensing of Viscous Solutions for Biomedical Applications

Cells cultured in three-dimensional scaffolds express a phenotype closer to in vivo cells than cells cultured in two-dimensional containers. Natural polymers are suitable materials to make three-dimensional scaffolds to develop disease models for high-throughput drug screening owing to their excellent biocompatibility. However, natural polymer solutions have a range of viscosities, and none of the currently available liquid dispensers are capable of dispensing highly viscous polymer solutions. Here, we report the development of an automated scaffold dispensing system for rapid, reliable, and homogeneous creation of scaffolds in well-plate formats. We employ computer-controlled solenoid valves to regulate air pressure impinging upon a syringe barrel filled with scaffold solution to be dispensed. Automated dispensing of scaffold solution is achieved via a programmable software interface that coordinates solution extrusion and the movement of a dispensing head. We show that our pneumatically actuated dispensing system can evenly distribute high-viscosity, chitosan-based polymer solutions into 96- and 384-well plates to yield highly uniform three-dimensional scaffolds after lyophilization. We provide a proof-of-concept demonstration of high-throughput drug screening by culturing glioblastoma cells in scaffolds and exposing them to temozolomide. This work introduces a device that can hasten the creation of three-dimensional cell scaffolds and their application to high-throughput testing.

High Throughput Genetic Characterisation of Caucasian Patients Affected by Multi-Drug Resistant Rheumatoid or Psoriatic Arthritis.

Rheumatoid and psoriatic arthritis (RA and PsA) are inflammatory rheumatic disorders characterised by a multifactorial etiology. To date, the genetic contributions to the disease onset, severity and drug response are not clearly defined, and despite the development of novel targeted therapies, ~10% of patients still display poor treatment responses. We characterised a selected cohort of eleven non-responder patients aiming to define the genetic contribution to drug resistance. An accurate clinical examination of the patients was coupled with several high-throughput genetic testing, including HLA typing, SNPs-array and Whole Exome Sequencing (WES). The analyses revealed that all the subjects carry very rare HLA phenotypes which contain HLA alleles associated with RA development (e.g., HLA-DRB1*04, DRB1*10:01 and DRB1*01). Additionally, six patients also carry PsA risk alleles (e.g., HLA-B*27:02 and B*38:01). WES analysis and SNPs-array revealed 23 damaging variants with 18 novel “drug-resistance” RA/PsA candidate genes. Eight patients carry likely pathogenic variants within common genes (CYP21A2, DVL1, PRKDC, ORAI1, UGT2B17, MSR1). Furthermore, “private” damaging variants were identified within 12 additional genes (WNT10A, ABCB7, SERPING1, GNRHR, NCAPD3, CLCF1, HACE1, NCAPD2, ESR1, SAMHD1, CYP27A1, CCDC88C). This multistep approach highlighted novel RA/PsA candidate genes and genotype-phenotype correlations potentially useful for clinicians in selecting the best therapeutic strategy.

The alamar blue assay in the context of safety testing of nanomaterials.

The Alamar Blue (AB) assay is widely used to investigate cytotoxicity, cell proliferation and cellular metabolic activity within different fields of toxicology. The use of the assay with nanomaterials (NMs) entails specific aspects including the potential interference of NMs with the test. The procedure of the AB assay applied for testing NMs is described in detail and step-by-step, from NM preparation, cell exposure, inclusion of interference controls, to the analysis and interpretation of the results. Provided that the proper procedure is followed, and relevant controls are included, the AB assay is a reliable and high throughput test to evaluate the cytotoxicity/proliferation/metabolic response of cells exposed to NMs.

Fluorimetric Assay of FAAH Activity.

Fatty acid amide hydrolase (FAAH) is the enzyme responsible for the degradation of anandamide (N-arachidonoylethanolamine, AEA) to arachidonic acid (AA) and ethanolamine. The method described here measures FAAH activity through the fluorometric arachidonoyl-7-amino-4-methyl-coumarin amide (AAMCA) substrate, which allows a simple and sensitive assay suitable for high-throughput screening tests. FAAH catalyzes the hydrolysis of AAMCA producing AA and the highly fluorescent compound 7-amino-4-methylcoumarin (AMC).

Embryo tracking system for high-throughput sequencing-based preimplantation genetic testing

Can the embryo tracking system (ETS) increase safety, efficacy and scalability of massively parallel sequencing-based preimplantation genetic testing (PGT)? Applying ETS-PGT, the chance of sample switching is decreased, while scalability and efficacy could easily be increased substantially. Although state-of-the-art sequencing-based PGT methods made a paradigm shift in PGT, they still require labor intensive library preparation steps that makes PGT cost prohibitive and poses risks of human errors. To increase the quality assurance, efficiency, robustness and throughput of the sequencing-based assays, barcoded DNA fragments have been used in several aspects of next-generation sequencing (NGS) approach. We developed an ETS that substantially alleviates the complexity of the current sequencing-based PGT. With (n = 693) and without (n = 192) ETS, the downstream PGT procedure was performed on both bulk DNA samples (n = 563) and whole-genome amplified (WGAed) few-cell DNA samples (n = 322). Subsequently, we compared full genome haplotype landscapes of both WGAed and bulk DNA samples containing ETS or no ETS. We have devised an ETS to track embryos right after whole-genome amplification (WGA) to full genome haplotype profiles. In this study, we recruited 322 WGAed DNA samples derived from IVF embryos as well as 563 bulk DNA isolated from peripheral blood of prospective parents. To determine possible interference of the ETS in the NGS-based PGT workflow, barcoded DNA fragments were added to DNA samples prior to library preparation and compared to samples without ETS. Coverages and variants were determined. Current PGT protocols are quality sensitive and prone to sample switching. To avoid sample switching and increase throughput of PGT by sequencing-based haplotyping, six control steps should be carried out manually and checked by a second person in a clinical setting. Here, we developed an ETS approach in which one step only in the entire PGT procedure needs the four-eyes principal. We demonstrate that ETS not only precludes error-prone manual checks but also has no effect on the genomic landscape of preimplantation embryos. Importantly, our approach increases efficacy and throughput of the state-of-the-art PGT methods. Even though the ETS simplified sequencing-based PGT by avoiding potential errors in six steps in the protocol, if the initial assignment is not performed correctly, it could lead to cross-contamination. However, this can be detected in silico following downstream ETS analysis. Although we demonstrated an approach to evaluate purity of the ETS fragment, it is recommended to perform a pre-PGT quality control assay of the ETS amplicons with non-human DNA, such that the purity of each ETS molecule can be determined prior to ETS-PGT. The ETS-PGT approach notably increases efficacy and scalability of PGT. ETS-PGT has broad applicative value, as it can be tailored to any single- and few-cell sequencing approach where the starting specimen is scarce, as opposed to other methods that require a large number of cells as the input. Moreover, ETS-PGT could easily be adapted to any sequencing-based diagnostic method, including PGT for structural rearrangements and aneuploidies by low-pass sequencing as well as non-invasive prenatal testing. M.Z.E. is supported by the EVA (Erfelijkheid Voortplanting & Aanleg) specialty program (grant no. KP111513) of Maastricht University Medical Centre (MUMC+), and the Horizon 2020 innovation (ERIN) (grant no. EU952516) of the European Commission. N/A.

High-throughput tensile testing of silver nanowires

Silver nanowires (Ag NWs) have been used in various nanostructured materials such as stretchable electronics due to their small size, high strength, and high conductivity. Envisioned nanostructured materials employ millions of nanostructures to achieve its function, yet due to challenges in sample preparation, mechanical characterization has been limited to a small number of samples. The results thus have limited statistical significance, important, for example, for the case of fracture strain, which is a stochastic variable. Here, we show highly-dense and unidirectional alignment of penta-twinned Ag NWs on top of polyethylene terephthalate (PET) substrates using microfluidics. Surface treatment enabled strong bonding between NWs and substrates. We then conducted tensile stretching of the substrates in the direction of alignment, capturing images of the induced deformation and fracture of the NWs, effectively testing many of them in tension simultaneously. We analyzed their fracture strain using digital image correlation (DIC). We show results for more than 200 samples, achieving a statistical quantification of NW fracture strain, useful for the design of applications and nanostructured materials based on them, such as stretchable electronics.

High-throughput and automated screening for COVID-19.

The COVID-19 pandemic has become a global challenge for the healthcare systems of many countries with 6 million people having lost their lives and 530 million more having tested positive for the virus. Robust testing and a comprehensive track and trace process for positive patients are essential for effective pandemic control, leading to high demand for diagnostic testing. In order to comply with demand and increase testing capacity worldwide, automated workflows have come into prominence as they enable high-throughput screening, faster processing, exclusion of human error, repeatability, reproducibility and diagnostic precision. The gold standard for COVID-19 testing so far has been RT-qPCR, however, different SARS-CoV-2 testing methods have been developed to be combined with high throughput testing to improve diagnosis. Case studies in China, Spain and the United Kingdom have been reviewed and automation has been proven to be promising for mass testing. Free and Open Source scientific and medical Hardware (FOSH) plays a vital role in this matter but there are some challenges to be overcome before automation can be fully implemented. This review discusses the importance of automated high-throughput testing, the different equipment available, the bottlenecks of its implementation and key selected case studies that due to their high effectiveness are already in use in hospitals and research centres.

Research Progress in High-Throughput Screening of CO2 Reduction Catalysts

The conversion and utilization of carbon dioxide (CO2) have dual significance for reducing carbon emissions and solving energy demand. Catalytic reduction of CO2 is a promising way to convert and utilize CO2. However, high-performance catalysts with excellent catalytic activity, selectivity and stability are currently lacking. High-throughput methods offer an effective way to screen high-performance CO2 reduction catalysts. Here, recent advances in high-throughput screening of electrocatalysts for CO2 reduction are reviewed. First, the mechanism of CO2 reduction reaction by electrocatalysis and potential catalyst candidates are introduced. Second, high-throughput computational methods developed to accelerate catalyst screening are presented, such as density functional theory and machine learning. Then, high-throughput experimental methods are outlined, including experimental design, high-throughput synthesis, in situ characterization and high-throughput testing. Finally, future directions of high-throughput screening of CO2 reduction electrocatalysts are outlooked. This review will be a valuable reference for future research on high-throughput screening of CO2 electrocatalysts.

A Protein Microarray-Based Respiratory Viral Antigen Testing Platform for COVID-19 Surveillance.

High-throughput and rapid screening testing is highly desirable to effectively combat the rapidly evolving COVID-19 pandemic co-presents with influenza and seasonal common cold epidemics. Here, we present a general workflow for iterative development and validation of an antibody-based microarray assay for the detection of a respiratory viral panel: (a) antibody screening to quickly identify optimal reagents and assay conditions, (b) immunofluorescence assay design including signal amplification for low viral titers, (c) assay characterization with recombinant proteins, inactivated viral samples and clinical samples, and (d) multiplexing to detect a panel of common respiratory viruses. Using RT-PCR-confirmed SARS-CoV-2 positive and negative pharyngeal swab samples, we demonstrated that the antibody microarray assay exhibited a clinical sensitivity and specificity of 77.2% and 100%, respectively, which are comparable to existing FDA-authorized antigen tests. Moreover, the microarray assay is correlated with RT-PCR cycle threshold (Ct) values and is particularly effective in identifying high viral titers. The multiplexed assay can selectively detect SARS-CoV-2 and influenza virus, which can be used to discriminate these viral infections that share similar symptoms. Such protein microarray technology is amenable for scale-up and automation and can be broadly applied as a both diagnostic and research tool.

Performance of High Throughput SARS-CoV-2 Antigen Testing Compared to Nucleic Acid Testing.

ObjectiveIndependent assessment of SARS-CoV-2 antigen (COV2Ag) tests remains important as varying performance between assays is common. We assessed the performance of a new high-throughput COV2Ag test compared to SARS-CoV-2 nucleic acid amplification tests (NAAT).MethodsA total of 347 nasopharyngeal samples collected from January to October 2021 were assessed by NAAT as part of standard-of-care testing (CDC LDT or GeneXpert System, Cepheid) and COV2Ag using the ADVIA Centaur CoV2Ag assay (Siemens Healthineers).ResultsAmong NAAT positive specimens we found 82.4% agreement and in NAAT negative specimens we found 97.3% agreement (overall agreement 85.6%). In symptomatic persons, COV2Ag agreed with NAAT 90.0% (n = 291), and in asymptomatic persons, 62.5% (n = 56). Agreement between positive NAAT and COV2Ag increased at lower cycle threshold (Ct) values.ConclusionThe COV2Ag assay exceeded the World Health Organization minimum performance requirements of ≥ 80% sensitivity and ≥ 97% specificity. The COV2Ag assay is helpful for large scale screening efforts due to high-throughput and reduced wait times.

A How to Guide: Clinical Population Test Development and Authorization of MALDI-ToF Mass Spectrometry-Based Screening Tests for Viral Infections.

Applying MALDI-ToF mass spectrometry as a clinical diagnostic test for viruses is different from that of bacteria, fungi and other micro-organisms. This is because the systems biology of viral infections, the size and chemical nature of specific viral proteins and the mass spectrometry biophysics of how they are quantitated are fundamentally different. The analytical challenges to overcome when developing a clinical MALDI-ToF mass spectrometry tests for a virus, particularly human pathogenic enveloped viruses, are sample enrichment, virus envelope disruption, optimal matrix formulation, optimal MALDI ToF MS performance and optimal spectral data processing/bioinformatics. Primarily, the instrument operating settings have to be optimized to match the nature of the viral specific proteins, which are not compatible with setting established when testing for bacterial and many other micro-organisms. The capacity to be a viral infection clinical diagnostic instrument often stretches current mass spectrometers to their operational design limits. Finally, all the associated procedures, from sample collection to data analytics, for the technique have to meet the legal and operational requirement for often high-throughput clinical testing. Given the newness of the technology, clinical MALDI ToF mass spectrometry does not fit in with standard criteria applied by regulatory authorities whereby numeric outputs are compared directly to similar technology tests that have already been authorized for use. Thus, CLIA laboratory developed test (LDT) criteria have to be applied. This article details our experience of developing a SAR-CoV-2 MALDI-ToF MS test suitable for asymptomatic carrier infection population screening.