Liquid System Research Articles

UV‐Triggered In Situ Formation of a Robust SEI on Black Phosphorus for Advanced Energy Storage: Boosting Efficiency and Safety via Rapid Charge Integration Plasticity

AbstractBlack phosphorus (BP) emerges as a highly promising electrode material for next generation of energy‐storage systems. Yet, its full potential is hindered by the instability of the solid‐electrolyte interphase (SEI) and the inflammability of its liquid systems. Here a pioneering UV‐induced in situ strategy is introduced for SEI construction, which leverages rapid electron supply to fracture sulfur‐dihalide bonds. This technique yields internal dihalide inorganic components and an external polymer segment, with any excess organic material being purged through pores. The (E)‐2‐chloro‐4‐((3′‐chloro‐4′‐hydroxyphenyl)diazinyl)phenyl acrylate (CA), with chlorine‐terminated groups, is initially in situ transformed into a flame‐retardant phenyl carboxylic acid (PCA), and then encapsulated within an ultrathin BP nanostructure, further nested in nitrogen (N), boron (B) co‐doped carbon (C) sheets that accommodate cobalt (Co) single atoms/nanoclusters (Co‐NBC). The Co‐NBC@BP@PCA construct demonstrates an impressive initial Coulombic efficiency (ICE) of 99.1% and maintains exceptional stabilities in terms of mechanical, chemical, and electrochemical performancecritical for prolonged cycle and calendar life. This research sheds light on the interplay between the rapid charge supply integrated in situ plasticity (RSIP) approach and the proactive establishment of an artificial SEI layer, offering profound insights into enhancing the durability and providing a solid foundation for advancements in energy storage technology.

Synergistic bioremediation of petroleum-contaminated soil using immobilized consortium of Rhodococcus rhodochrous and Bacillus subtilis laccase

Petroleum-contaminated soil represents a significant environmental and public health challenge on a global scale. Microbial bioremediation has shown potential, yet the role of enzymes in enhancing petroleum degradation remains underexplored. In this study, the synergistic effects of Rhodococcus rhodochrous (R.rh) and Bacillus subtilis-derived laccase (BsLac) was investigated in the remediation of petroleum-contaminated soil. Immobilized R.rh (PSIMRH) and BsLac (ADIMLac) exhibited higher petroleum degradation rates than their free state, achieving 78.3% and 56.3% degradation in liquid systems, respectively. The combined treatment of PSIMRH and ADIMLac demonstrated a synergistic effect on petroleum degradation, achieving 43.6% with a maximum degradation constant of 0.0335 d−1, representing a 202.7% improvement over untreated soil. PSIMRH enhanced petroleum degradation through microbial metabolism, while ADIMLac accelerated the initial breakdown of complex hydrocarbons into simpler, more bioavailable ones via enzymatic oxidation, providing growth substrates for microbes and significantly improving petroleum degradation rates. The microbial analysis revealed an increase abundance of known petroleum-degrading bacterial genera, including Rhodococcus, Lysobacter, Micromonospora, and Streptomyces. However, the presence of BsLac appeared to reduce the competitive advantage of Rhodococcus, promoting the proliferation of indigenous strains like Lysobacter and Streptomyces. These results suggest that enzyme-microbe synergy can enhance the bioremediation process by altering microbial community dynamics and accelerating petroleum degradation. This study attempts to remediate petroleum-contaminated pollution with the combined use of strains and enzymes, providing a new approach for the remediation of other pollution problems.

Numerical Analysis of Optimising Liquid Desiccant Dehumidification for Sustainable Building Cooling: A Data-Driven Method Using Response Surface Methodology

Leveraging data-driven methods such as Response Surface Methodology (RSM) has considerable potential for sustainable building cooling via mitigating energy consumption and environmental impacts. This research focuses on using the RSM to improve liquid desiccant dehumidification for sustainable building cooling performance using a D-optimal design. Specifically, the research intends to investigate the actual influence of the inlet air conditions and desiccant concentration on the performance of liquid desiccant dehumidification systems, i.e., the moisture removal rates and dehumidifier efficiency. To systematically conduct this research, a set of experimental data gathered from the open literature is utilised. This includes a specific set of inlet parameters of air temperature (27–34.5 °C), ratio of air humidity (20.5–25 g/kg), and solution temperature (27.5–38.5 °C) as the independent variables. Also, the feedback variables include the moisture removal rates (MRR) and efficacy (ϵ). The associated results of the analysis of variation indicate that the ratio of air humidity has the greatest influence on the moisture removal rate. However, the solution temperature and the ratio of air humidity have the most influence on efficacy. In the event of response optimisation, the result at MRR and (ϵ) are 0.54 g/s and 0.50, respectively, with a minimum desirability of 0.992 and 1.

Case-control study on associations of hemorrhagic bowel syndrome in swine with feed characteristics and intestinal pathogens

Haemorrhagic bowel syndrome (HBS) is one of the most common causes of death in fattening pigs worldwide. The objective of this descriptive study was to systematically assess predictors or causal components for the appearance of HBS using case farms (mortality rate caused by HBS ≥ 1.5%) in comparison with control farms (mortality rate caused by HBS ≤ 0.25%), focusing on feed ingredients, feed quality and size, and gastrointestinal pathogens. The inclusion of sugar beet as a feed component in liquid feeding systems was found to be associated (p = 0.03) with farms identified as HBS cases. Another predictive or causal factor found for liquid feeding systems, but only for those using meal, was particle size. A higher percentage of small particles (< 2 mm) in the meal was associated with a higher risk of being an HBS case farm (p = 0.02), while no relevant association was detected for the use of pellets. Sugar beet in the diet was also associated with the incidence of HBS.The microbial quality of the feed in dry feeding systems, specifically the number of total aerobes at the first and last outlet tubes, was associated with a higher incidence of HBS (p = 0.03). Faecal sample analysis showed a difference (p < 0.05) in the prevalence of B. pilosicoli in the herd category (case vs. control herd). In this descriptive study, five predictive or causal factors were identified for an HBS farm with a mortality rate due to HBS ≥ 1.5%. These included the number of aerobes in dry matter samples from the first and last feeders, the particle diameter of the meal used in liquid feeding systems and sugar beet as a component of liquid feeding rations, and the presence of B. pilosicoli as an infectious agent at animal level. Relevant associations reinforce the findings of the previously published Swiss study that HBS is a multifactorial syndrome involving different aspects of pig production and cannot be attributed to a single cause. Further studies are needed to develop evidenced based causal models for HBS in swine.

Innovations in Nanoemulsion Technology: Enhancing Drug Delivery for Oral, Parenteral, and Ophthalmic Applications.

Nanoemulsions (NEs) are submicron-sized heterogeneous biphasic liquid systems stabilized by surfactants. They are physically transparent or translucent, optically isotropic, and kinetically stable, with droplet sizes ranging from 20 to 500 nm. Their unique properties, such as high surface area, small droplet size, enhanced bioavailability, excellent physical stability, and rapid digestibility, make them ideal for encapsulating various active substances. This review focuses on recent advancements, future prospects, and challenges in the field of NEs, particularly in oral, parenteral, and ophthalmic delivery. It also discusses recent clinical trials and patents. Different types of in vitro and in vivo NE characterization techniques are summarized. High-energy and low-energy preparation methods are briefly described with diagrams. Formulation considerations and commonly used excipients for oral, ocular, and ophthalmic drug delivery are presented. The review emphasizes the need for new functional excipients to improve the permeation of large molecular weight unstable proteins, oligonucleotides, and hydrophilic drugs to advance drug delivery rapidly.

Establishment, Multiplication, and Biochemical Analysis of Embryogenic Lines of the Amazonian Palm Euterpe precatoria Mart. under Suspension Culture

The palm Euterpe precatoria holds great social, cultural, and environmental importance. The heart of palm and the fruit are the main products used for industrialization due to their energetic properties. Thus, the aim of this study was to establish a suspension cultivation protocol for the species using different explant sources. For this, eight lineages of E. precatoria embryogenic calluses were tested, with five in liquid medium Murashige and Skoog (MS) with 5 μM Picloram and three for comparison in semisolid medium MS with 20 μM Picloram and 5 μM 2iP. The growth curve was obtained by weighing the calli from 60 to 180 days of cultivation. The Gompertz model was applied, and growth kinetics were evaluated. At 100 days, the contents of total soluble sugars (TSSs) and total soluble proteins (TSPs) were determined. Principal components (PCA) were measured. According to the analysis of the data, the cultivation of E. precatoria lineages in liquid medium was successfully carried out, and the establishment was achieved. The model can be considered adequate since the R2 values found describe more than 90% of the growth kinetics of the lineages. In the liquid system, lineages L1 (from leaf explants and multiplied in semisolid medium—SM), L2 (from leaf explants and multiplied in SM), and L6 (from zygotic embryo explants and multiplied in liquid medium—LM) showed the shortest time to double the biomass accumulation. Multivariate analysis reveals a significant increase in masses in liquid cultures, represented by lineages L6 and L2. There was statistical difference in the amount of TSSs extracted; the highest TSS levels were observed in lineages cultivated in LM. The protein content found was very low, showing statistical differences among the lineages. In this work, the establishment and multiplication of embryogenic calli of E. precatoria are described for the first time, and they emerge as viable alternatives for the vegetative propagation of the species.

Numerical simulation of fluid flow characteristics and ultrasonic cavitation performance in novel-designed stirred tank sonoreactor

A novel-designed stirred tank sonoreactor was developed to well solve large-scale dispersion of nanoparticles in liquid phase system. The fluid flow characteristics and ultrasonic cavitation performance in the sonoreactor were numerically simulated by CFD method. By comparing the pressure, cavitation bubble and velocity distribution under different situations, it is found that larger ultrasonic amplitude, relatively smaller ultrasonic frequency, higher saturated vapor pressure and lower viscosity of liquid medium are beneficial to ultrasonic cavitation. Besides, the acoustic flow action is strengthened with the increase of ultrasonic frequency and decrease of gas-liquid mixture's average density. For the designed sonoreactor, it is critical that high-frequency transducers should be determined near the bottom and upper region of the kettle, and large-amplitude transducers can be confirmed in the middle position. The research findings will provide theoretical and technical supports for developing state-of-the-art sonoreactors and optimizing ultrasonic process parameters in the industrialized preparation of nanocomposites.

Poly(lactide) Upcycling Approach through Transesterification for Stereolithography 3D Printing.

The legislature determines the recycled and waste contents in fabrication processes to ensure more sustainable production. PLA's mechanical recycling and reuse are limited due to the performance decrease caused by thermal or hydrolytic instability. Our concept introduces an upcycling route involving PLA depolymerization using propylene glycol as a reactant, followed by the methacrylation, assuring the liquid systems' curability provided by radical polymerization. PLA-containing curable systems were studied from a rheological and thermomechanical viewpoint. The viscosity levels varied from 33 to 3911 mPa·s at 30 °C, giving a wide capability potential. The best system reached 2240 MPa storage modulus, 164.1 °C glass-transition temperature, and 145.6 °C heat-resistant index, competitive values to commercial systems. The printability was verified for all of the systems. Eventually, our concept led to SLA resin production containing PLA waste content up to 51 wt %.

Affordable and Reliable RP-HPLC Method for Verapamil Hydrochloride Quantification in Rabbit Plasma for Pharmacokinetics

Background: Existing bioanalytical methods for verapamil hydrochloride (VH) are often complex, requiring advanced instrumentation and specialized expertise, which limits their use in resource-constrained laboratories. Aim: The goal of this study is to fill this gap by developing a simplified, robust RP-HPLC-UV approach for the estimation of verapamil hydrochloride in rabbit plasma. Designed to enhance accuracy and precision while minimizing sample preparation challenges, this method addresses existing limitations by providing an affordable and reliable alternative for laboratories lacking sophisticated instrumentation. Methods: The bioanalytical method was implemented on C-18 stationary phase (5 μ, 250 × 4.6 mm) using acetonitrile/0.1% tetrahydrofuran (THF) in water (80:20, in volume) as the liquid system at a 1 mL/min flow speed, employing carvedilol as an internal standard. Results: The reported retention times of verapamil hydrochloride and carvedilol were ~7.64 and 4.69 min, respectively, at sufficiently high system suitability standards. The linearity of the bioanalytical approach can be seen between 0.025 and 5.0 µg/mL (r2 = 0.9991). The findings indicated that there was no matrix influence in terms of accuracy (≥98.96 ± 2.68%), intra- and inter-day precision (≤3.68%), recovery (101.98 ± 2.76%), and procedure efficiency (100.65 ± 1.82%). Benchtop, long-term, and short-term stability investigations all revealed that the verapamil hydrochloride in the bio-samples was stable. The pharmacokinetic parameters (Cmax—3.47 µg/mL; Tmax—1.59 h) were studied from time-dependent plasma concentrations of verapamil hydrochloride estimated after 40 mg oral dosing in New Zealand white rabbits. Conclusions: The developed bioanalytical method provided easier quantitative analysis of verapamil hydrochloride from rabbit plasma and was effectively used in a pharmacokinetic investigation of an oral bolus. The reliable performance of this method under practical conditions positions it as a crucial tool for advancing pharmacokinetic studies across various research environments.

Evaluation of microplate handling accuracy for applying robotic arms in laboratory automation

An inexpensive single-arm robot is widely utilized for recent laboratory automation solutions. The integration of a single-arm robot as a transfer system into a semi-automatic liquid dispenser without a transfer system can be realized as an inexpensive alternative to a fully automated liquid handling system. However, there has been no quantitative investigation of the positional accuracy of robot arms required to transfer microplates. In this study, we constructed a platform comprising aluminum frames and digital gauges to facilitate such measurements. We measured the position repeatability of a robot arm equipped with a custom-made finger by repeatedly transferring microplates. Further, the acceptable misalignment of plate transfer was evaluated by adding an artificial offset to the microplate position using this platform. The results of these experiments are expected to serve as benchmarks for the selection of robot arms for laboratory automation in biology. Furthermore, all information for replicating this device will be made publicly available, thereby allowing many researchers to collaborate and accumulate knowledge, hopefully contributing to advances in this field.

Analysis of molecular interactions of an aqueous benzoates with glycolic and lactic acid: spectroscopic, acoustic, and volumetric approach

Volumetric as well as Acoustic properties for the ternary liquid combination (water + sodium methyl p-hydroxybenzoate + glycolic acid/lactic acid) are evaluated at several temperatures using a Densimeter (DSA 5000 M) over the range of concentrations (0.000, 0.015, 0.025, 0.035) mol.kg−1. Several thermo-acoustical properties were computed utilizing the experimental data. These values were then used to derive several parameters, including the apparent molar volume( V∅ ); partial molar volume (V∅0); apparent molar isentropic compression (K∅,S); transfer properties(Δ V∅0, Δ K∅,s0 ); partial molar isentropic compression (K∅,s0); apparent molar expansibility ( E∅0 ) as well as its first derivative ( ∂Eϕ0/∂T ) P; and coefficient of thermal expansivity( αp ). The obtained results are explained by analyzing the several interactions that take place inside the liquid system by implementing the theory of co-sphere overlap. Utilizing these thermodynamical parameters, the interaction coefficients are computed to evaluate the ternary mixture corresponding to the interactions between solutes and solvents. FTIR spectrum analysis revealed the presence of H-bonds along with the other molecular interactions in the analyzed systems, as shown by the shifting of the O-H stretching band.

Novel ionic liquid systems based on three-nitro phenoxide: Spectroscopic and electronic characterization using theoretical and experimental study

After extensive computations, novel trinitro compounds, triethyl ammonium 2,4,6-trinitro phenoxide (TEA-TNP) and tetramethyl guanidinium 2,4,6-trinitro phenoxide (TMG-TNP), are designed, synthesized, and identified. The Geometrical parameters, electronic properties, and spectroscopic analysis have been investigated by comparison of theoretical and experimental methods. Spectrum features that include IR and Raman have been calculated, and 1H and 13C NMR spectra are compared to experimental data. Electronic analysis has been investigated through NLO, FMO, UV-Vis, NBO, and MEP calculations. B3LYP/6-311++G(d,p) is highly successful and dependable for predicting ionic liquid stability, electrical characteristics, and spectroscopy. Both TEA-TNP and TMG-TNP are introduced as suitable candidates for NLO material. TMG-TNP with greater nucleophilicity (N) is a noncorrosive lubricant additive that reduces friction and wear for tribological performance.

Solid–liquid separation state control system for shale shaker based on computer vision

Shale shakers are crucial in drilling operations, separating solid–liquid mixtures, protecting equipment, improving efficiency, and supporting environmental protection and resource recovery. Typically, the solid–liquid separation status on shaker screens is assessed through manual inspection and experiential judgment, with the screen tilt angle adjusted manually. This method has inherent delays, failing to promptly detect and address screen surface abnormalities. To overcome this, a computer vision-driven intelligent control system has been proposed, enhancing drilling sieving operations’ efficiency. The system uses computer vision and servo motors to monitor and adjust the sieving state. Twin industrial cameras capture images, which undergo preprocessing like perspective correction, grayscale conversion, noise reduction, and illumination equalization. Local threshold segmentation and morphological analysis distinguish between liquid and solid states. The segmented images’ contours and area analysis precisely determine the separation state. Results are transmitted to motors via an industrial computer to adjust the sieve net angle. Servo motors on both sides manipulate the sliding block lifting mechanism for precise tilt adjustment. Experiments demonstrate that the method achieves an average accuracy of 93.635% in identifying mud edges. The system’s high efficiency and reliability ensure accurate identification and adjustment of the solid–liquid separation state, thereby optimizing drilling sieving workflows.

Valorization of Eggshell as Renewable Materials for Sustainable Biocomposite Adsorbents—An Overview

The production and buildup of eggshell waste represents a challenge and an opportunity. The challenge is that uncontrolled disposal of generated eggshell waste relates to a sustainability concern for the environment. The opportunity relates to utilization of this biomass resource via recycling for waste valorization, cleaner production, and development of a circular economy. This review explores the development of eggshell powder (ESP) from eggshell waste and a coverage of various ESP composite sorbents with an emphasis on their potential utility as adsorbent materials for model pollutants in solid–liquid systems. An overview of literature since 2014 outlines the development of eggshell powder (ESP) and ESP composite adsorbents for solid–liquid adsorption processes. The isolation and treatment of ESP in its pristine or modified forms by various thermal or chemical treatments, along with the preparation of ESP biocomposites is described. An overview of the physico-chemical characterization of ESP and its biocomposites include an assessment of the adsorption properties with various model pollutants (cations, anions, and organic dyes). A coverage of equilibrium and kinetic adsorption isotherm models is provided, along with relevant thermodynamic parameters that govern the adsorption process for ESP-based adsorbents. This review reveals that ESP biocomposite adsorbents represent an emerging class of sustainable materials with tailored properties via modular synthetic strategies. This review will serve to encourage the recycling and utilization of eggshell biomass waste and its valorization as potential adsorbent systems. The impact of such ESP biosorbents cover a diverse range of adsorption-based applications from environmental remediation to slow-release fertilizer carrier systems in agricultural production.

Liquid Handling Technologies: A Study through Major Discoveries and Advancements

Liquid handling is an essential process in laboratories involving automatically transferring accurate amounts of liquids for various task completions such as sample preparation, pipetting, dilution, and mixing. Automated liquid handling systems (ALHS) help to minimize the errors, complications, and difficulties that are faced with manual methods of liquid handling by efficiently enhancing the accuracy, precision, and reproducibility of experimental workflows and have transformed life sciences aiding research in various fields. Processes like Nuclear Magnetic Resonance (NMR) spectrometry, mass spectrometry, and acoustic droplet ejection with ALHS showcase tremendous potential. Outdated manual machinery is being replaced rapidly with automated tools. We have observed that these laboratory setups are sophisticated and costly. It is challenging to access such technologies, especially in poor or developing countries. At the nanoscale level, handling and evaporation of biomaterials becomes a great issue. Multitudinous devices can combat the cost issue along with utilization of single equipment without the enhancement of add-on modules. Liquid handling devices enhance the quality of experiments contributing to efficient dealing of circumstances like COVID-19. They aid in genomics and proteomics and play a pivotal role in liquid manipulation, drug screening, quantifications, forensic studies and many more. Liquid handling devices with robotic equipment can perform multiple tasks and can assist in a wide range of fields. We anticipate that in the coming years, our laboratories will be furnished with advanced, robotic technologies and cause a revolution in the scientific scenery.

A subchannel analysis code for advanced liquid metal fast reactor cores and study on heat transfer characteristics of core geometry parameters

The liquid metal fast reactor represents an important reactor type for the future development of nuclear energy. Accurately modeling and predicting the subchannel flow and heat transfer phenomenon in the rod bundles plays a key role in ensuring core safety. Consequently, a subchannel code suitable for the liquid metal fast reactor is analyzed based on the subchannel analysis code of the Pressurized Water Reactor (PWR). The modified code incorporates various types of liquid metals, such as lead, lead-bismuth eutectic (LBE), and sodium, along with their constitutive models, enhancing the applicability of the liquid metal reactor systems. This code has been verified and validated at several levels, including comparing the code simulation results with other similar subchannel codes results, high-dimensional Computational Fluid Dynamics (CFD) simulations, and experiment data. The sensitivity analysis of core geometric parameters and turbulent mixing coefficients is performed based on the modified version code. The influence of the core geometry, including the rod Pitch to Diameter ratio (P/D), Rod to Wall gap (RTW) and the wire wrap pitch (H) on the sensitivity of outlet temperature has been investigated. The results show that the new subchannel analysis code suitable for liquid metal cooled reactor shows reasonable agreement with the verified and validated results. The geometric parameters, such as P/D, RTW and H have noticeable effects on the outlet temperature difference of the subchannels. Additionally, the outlet temperature distribution in the subchannel is significantly affected by different turbulent mixing coefficients and the distribution of the turbulent mixing coefficient also influenced by the reactor core sizes. Overall, this study could support the rod bundles design and safety analysis in the liquid metal reactors.

Optimization of a novel liquid carbon dioxide energy storage system by thermodynamic analysis and use of solar energy and liquefied natural gas

Liquid carbon dioxide energy storage with its advantages in terms of geographical constraints and economic performance has garnered significant attention. In this study, a novel liquid carbon dioxide storage system was proposed which utilizes the waste cold energy from LNG and achieves high liquefaction efficiency. By integrating solar energy, net output of the system is increased. The results show that the net output is sensitive to turbine 1 outlet pressure. Round-trip efficiency and energy storage density increases with the rise of the pump outlet pressure first and subsequently exhibits a gradual decline with an inflection point. With the increase in ambient temperature, the system efficiency shows a slight decrease, yet energy storage density shows a significant increase. At basic operating conditions, the system demonstrates energy storage efficiency of 66.64 %, round-trip efficiency of 74.33 %, and energy storage density of 23.51 kWh/m3. In addition, optimization was conducted. Single-objective optimization results show that round-trip efficiency, energy storage density and levelized cost of storage can reach 89.86 %, 25.37 kWh/m3 and 0.1873 $/kWh, respectively. Multi-objective optimization obtained compromise results of round-trip efficiency and levelized cost of storage of 85.33 % and 0.1947 $/kWh when integrated solar energy, showing a 33.77 % increase and a 13.20 % decrease compared to 63.79 % and 0.2243 $/kWh when not integrated solar energy.

The Degradation of the 2-Hydroxyethylhydrazinium Nitrate Ionic Liquid System.

The mechanism by which the 2-hydroxyethylhydrazinium nitrate (HEHN) ionic liquid (IL) dissociates via the formation and loss of nitric acid is investigated utilizing the effective fragment potential (EFP) method and various ab initio methods. Additionally, the interactions that dictate the stability of the HEHN IL and contribute to the formation of nitric acid are further assessed using the quasi-atomic orbital (QUAO) bonding analysis. From this work, it is suggested that the formation of nitric acid is dictated by the presence of hydrogen-bonding interactions which appear to become increasingly ionic in nature as the system approaches the bulk, limiting the formation of nitric acid.

B-025 Revolutionizing NGS-driven Laboratory Developed Tests: The impact of automated liquid handling on plate-based bead clean-up

Abstract Background Automated liquid handling systems are proving to be a transformative technology in the landscape of Laboratory Developed Tests (LDTs), especially those utilizing Next-Generation Sequencing (NGS). By enhancing accuracy, boosting efficiency, and offering scalable solutions, these systems enable labs to meet current and future challenges more effectively. The right system not only improves the quality of diagnostics but also positions labs to be agile and adaptive in a rapidly advancing healthcare environment. firefly® is a well-suited liquid handling solution for diagnostics labs developing LDTs due to its compact footprint, user-friendly software and exceptional versatility. Validated protocol templates that support NGS library preparation are readily available from the firefly cloud and can be adapted to meet specific lab needs. For example, automated bead-based clean-ups designed for 96- and 384- NGS applications are available on firefly. These remove DNA fragments of undesirable sizes, enzymes or primers rom the amplified DNA after a PCR reaction. Automating this process facilitates higher throughput and scaling. firefly has a unique combination of 6 positive displacement dispense heads and a 96/384-well air-displacement pipetting head, making it ideal to distribute the reagents required for bead clean-ups. Methods firefly protocols were developed and optimized to be compatible with a range of elution volumes and bead ratios. Protocol performance was tested using library DNA (∼300bp). The same input DNA was plated out and quantified using the QuantiFluor dsDNA system (Promega) before and after running the protocol. This data was used to assess any reduction in DNA concentration and uniformity across the plate. The protocol's ability to successfully modulate the fragment size distribution was demonstrated using a 50bp DNA ladder and different bead ratios. Data was obtained using a Fragment Analyzer (Agilent, DNF-474 HS NGS Fragment Kit). Well-to-well contamination was assessed by running no-template-controls (NTCs) in wells adjacent to library DNA, then analyzed by qPCR using a LightCycler 480 System (Roche, KAPA Library Quantification kit). Results 96 format has a setup time of 10 minutes, run time of 43 and %CV library concentration of &amp;lt;6%. 384 format has a setup time of 10 minutes, run time of 47 and %CV library concentration of &amp;lt;8%. Uniformity of concentration and fragment sizes was seen across the plates. Comparison of the library concentration before and after the clean-up showed a 5% reduction in DNA concentration after the 96-well plate clean-up and an 11% reduction in library concentration when using the 384-well plate clean-up. Fragment size analysis showed that fragment sizes can be successfully modulated on firefly by changing the bead ratio of the firefly run and no detectable well-to-well contamination was detected by qPCR. Conclusions We have demonstrated that automated bead clean-ups on firefly are uniform show uniformity across both 96- and 384-well plates, with no detectable well-to-well contamination, to deliver the consistent and robust results required for an LDT. Bead-based cleanups on firefly offer a fast, 100% walkaway solution. By running 96-samples at once on firefly, we see approximately a 6-fold increase in throughput than when performed manually by a single user.

Enhancing quantitative imaging to study DNA damage response: A guide to automated liquid handling and imaging

Laboratory automation and quantitative high-content imaging are pivotal in advancing diverse scientific fields. These innovative techniques alleviate the burden of manual labour, facilitating large-scale experiments characterized by exceptional reproducibility. Nonetheless, the seamless integration of such systems continues to pose a constant challenge in many laboratories. Here, we present a meticulously designed workflow that automates the immunofluorescence staining process, coupled with quantitative high-content imaging to study DNA damage signalling as an example. This is achieved by using an automatic liquid handling system for sample preparation. Additionally, we also offer practical recommendations aimed at ensuring the reproducibility and scalability of experimental outcomes. We illustrate the high level of efficiency and reproducibility achieved through the implementation of the liquid handling system but also addresses the associated challenges. Furthermore, we extend the discussion into critical aspects such as microscope selection, optimal objective choices, and considerations for high-content image acquisition. Our study streamlines the image analysis process, offering valuable recommendations for efficient computing resources and the integration of cutting-edge deep learning techniques. Emphasizing the paramount importance of robust data management systems aligned with the FAIR data principles, we provide practical insights into suitable storage options and effective data visualization techniques. Together, our work serves as a comprehensive guide for life science laboratories seeking to elevate their high-content quantitative imaging capabilities through the seamless integration of advanced laboratory automation.