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Commercial automation systems for small- and medium-sized laboratories, including research environments, are often complex to use. For liquid handling systems (LHS), development is required not only for the robot's movements but also for adapting the bioanalytical method to the automated system. This study investigates whether a more human-like automation strategy—using a robotic system (RS)—is more suitable for research laboratories than a professional automation approach utilizing a commercial automated LHS. We conducted a series of measurements for protein determination using a Bradford assay manually, with a fully automated LHS, and with our human-like RS. Although the hand-like RS approach requires more than twice the time of the LHS, it achieved the best standard deviation in this setup (RS = 0.5, manual = 0.71, LHS = 0.86). Due to the low limit of detection (LOD) and limit of quantification (LOQ), most protein samples could be quantified with the RS (samples below LOQ = 9.7%, LOD = 0.23; LOQ = 0.25) compared to manual (samples below LOQ = 28.8%, LOD = 0.24; LOQ = 0.26) and the LHS (samples below LOQ = 36.1%, LOD = 0.27; LOQ = 0.31). In another time-dependent enzymatic assay test, the RS achieved results comparable to the manual method and the LHS, although the required time could be a constraint for short incubation times. Our results demonstrate that a more hand-like automation system closely models the manual process, leading easier to accurate bioanalytical results. We conclude that such a system could be more suitable for typical research environments than a complex LHS. [ABSTRACT FROM AUTHOR]

The implementation of automation has already had a considerable impact on chemical and pharmaceutical industrial laboratories. However, academic laboratories have often been more reluctant to adopt such technology due to the high cost of commercial liquid handling systems, although, in many instances, there would be a huge potential to automate repetitive tasks, resulting in elevated productivity. We present here a detailed description of the setup, validation, and utilization of a multifunctional liquid automation (MULA) system that can be used to automate various chemical and biological tasks. Considering that such a setup must be highly customizable, we also designed MULA with respect to modularity, providing detailed insight as far as possible. Including all 3D-printed parts and the used Hamilton gastight micro syringe, the total construction cost is approximately 700 €. This allows us to achieve a highly reliable and accurate system that exceeds the precision of a classical air displacement pipette while still retaining the ability to use closed vial (septa) setups. To encourage other groups to adopt this setup, detailed instructions and tips for every step of the process are provided, along with the complete CAD design of MULA and control code, which are freely available for download under the CC BY NC 3.0 license.

Recent advances in the development of automation hardware, such as liquid handling robots, offer accelerated and less labor-intensive workflows for material synthesis and diagnostic assays. Furthermore, greatly increased data redundancy and removal of the experimenter variability are expected to contribute to higher reliability and reproducibility of communicated results. One current limitation for further deployment, especially in open hardware solutions, is the integration of centrifuges in automated workflows. Conventional centrifuges are often not suitable for portable or modular applications and, crucially, do not offer the required positional control over the samples. This work presents a centrifuge with infrared sensor-derived positional control based on open-source design and cost-effective manufacturing. Users can accurately control the centrifugation parameters via a serial port. Besides, this device is built with modular components, allowing for further customized adaptations. The manuscript presents the design, construction, and general validation, as well as the functionality testing and quality control. By using an adaptive bottom plate, the smart centrifuge is compatible with various deck configurations, here demonstrated for the open-source pipetting robot OT-2 (Opentrons). This accessible platform offers flexible use in automated synthesis and sample processing across various research fields in life sciences, physical sciences, and engineering.

Polymerase chain reaction (PCR)-based assays were widely deployed during the SARS-CoV-2 pandemic for population-scale testing. High-throughput molecular diagnostic laboratories required a high degree of process automation to cope with huge testing demands, fast turnaround times, and quality requirements. However, process developers and optimizers often neglected the critical step of preparing a PCR Master Mix. The construction of PCR Master Mix depends on operator skill during the manual pipetting of reagents. Manual procedures introduce variation, inconsistency, wastage, and potentially risks data integrity. To address this, we developed a liquid-handler-based solution for automated, traceable, and compliant PCR Master Mix preparation. Here, we show that a fully automated PCR Master Mix protocol can replace manual pipetting, even in a diagnostic environment, without affecting accuracy or precision. Ultimately, this method eliminated operator-induced wastage and improved the consistency of the quality of results.

Column chromatography and the subsequent analysis of the collected fractions using thin-layer chromatography (TLC) are immensely popular and frequently used methods in organic chemistry. To analyze the fractions, they have to be spotted onto the TLC plates, which can be a time-consuming task. Therefore, we have developed an automatic spotting device based on a conversion of a 3D printer kit, which could be easily built for around 500 €. All parts come directly from the 3D printer kit, are 3D printed, or are commercially available, such as syringes and tubing. In this manuscript, we describe the design and use of the spotting device. In addition, we provide a detailed step-by-step assembly instruction and all CAD files for easy replication.

This study examines the intricate relationship between protein glycosylation dynamics and therapeutic responses in Luminal A and Luminal B breast cancer subtypes, focusing on anastrozole and tamoxifen impacts. The present methods inadequately monitor and forecast patient reactions to these treatments, leaving individuals vulnerable to the potential adverse effects of these medications. This research investigated glycan structural changes by following patients for up to 9 months. The protocol involved a series of automated steps including IgG isolation, protein denaturation, glycan labelling, purification, and final analysis using capillary gel electrophoresis with laser-induced fluorescence. The results suggested the significant role of glycan modifications in breast cancer progression, revealing distinctive trends in how anastrozole and tamoxifen elicit varied responses. The findings indicate anastrozole's association with reduced sialylation and increased core fucosylation, while tamoxifen correlated with increased sialylation and decreased core fucosylation. These observations suggest potential immunomodulatory effects: anastrozole possibly reducing inflammation and tamoxifen impacting immune-mediated cytotoxicity. This study strongly emphasizes the importance of considering specific glycan traits to comprehend the dynamic mechanisms driving breast cancer progression and the effects of targeted therapies. The nuanced differences observed in glycan modifications between these two treatments underscore the necessity for further comprehensive research aimed at thoroughly evaluating the long-term implications and therapeutic efficacy for breast cancer patients. [ABSTRACT FROM AUTHOR]

The large volumes of samples to be analysed every day would be impossible to manage without laboratory automation. As laboratory procedures have progressed, so have the tasks of laboratory personnel. With this feature article, we would like to provide (bio)chemical practitioners with little or no knowledge of laboratory automation with a guide to help them decide whether to implement laboratory automation and find a suitable system. Especially in small- and medium-sized laboratories, operating a laboratory system means having bioanalytical knowledge, but also being familiar with the technical aspects. However, time, budget and personnel limitations allow little opportunity for personnel to get into the depths of laboratory automation. This includes not only the operation, but also the decision to purchase an automation system. Hasty investments do not only result in slow or non-existent cost recovery, but also occupy valuable laboratory space. We have structured the article as a decision tree, so readers can selectively read chapters that apply to their individual situation. This flexible approach allows each reader to create a personal reading flow tailored to their specific needs. We tried to address a variety of perspectives on the topic, including people who are either supportive or sceptical of laboratory automation, personnel who want or need to automate specific processes, those who are unsure whether to automate and those who are interested in automation but do not know which areas to prioritize. We also help to make a decision whether to reactivate or discard already existing and unused laboratory equipment. [ABSTRACT FROM AUTHOR]

Every year biotechnology labs generate a combined total of ∼5.5 million tons of plastic waste. As the global bioeconomy expands, biofoundries will inevitably increase plastic consumption in-step with synthetic biology scaling. Decontamination and reuse of single-use plastics could increase sustainability and reduce recurring costs of biological research. However, throughput and variable cleaning quality make manual decontamination impractical in most instances. Automating single-use plastic cleaning with liquid handling robots makes decontamination more practical by offering higher throughput and consistent cleaning quality. However, open-source, validated protocols using low-cost lab robotics for effective decontamination of plasticware—facilitating safe reuse—have not yet been developed. Here we introduce and validate TidyTron: a library of protocols for cleaning micropipette tips and microtiter plates that are contaminated with DNA, E. coli, and S. cerevisiae. We tested a variety of cleaning solutions, contact times, and agitation methods with the aim of minimizing time and cost, while maximizing cleaning stringency and sustainability. We tested and validated these cleaning procedures by comparing fresh (first-time usage) versus cleaned tips and plates for contamination with cells, DNA, or cleaning solutions. We assessed contamination by measuring colony forming units by plating, PCR efficiency and DNA concentration by qPCR, and event counts and debris by flow cytometry. Open source cleaning protocols are available at https://github.com/PlantSynBioLab/TidyTron and hosted on a graphical user interface at https://jbryantvt.github.io/TidyTron/.

The Droplet Microarray (DMA) has emerged as a tool for high-throughput biological and chemical applications by enabling miniaturization and parallelization of experimental processes. Due to its ability to hold hundreds of nanoliter droplets, the DMA enables simple screening and analysis of samples such as cells and biomolecules. However, handling of nanoliter volumes poses a challenge, as manual recovery of nanoliter volumes is not feasible, and traditional laboratory equipment is not suited to work with such low volumes, and small array formats. To tackle this challenge, we developed the Automated Nanoliter Droplet Selection device (ANDeS), a robotic system for automated collection and transfer of nanoliter samples from DMA.ANDeS can automatically collect volumes from 50 to 350 nL from the flat surface of DMA with a movement accuracy of ±30 µm using fused silica capillaries. The system can automatically collect and transfer the droplets from DMA chip into other platforms, such as microtiter plates, conical tubes or another DMA. In addition, to ensure high throughput and multiple droplet collection, the uptake of multiple droplets within a single capillary, separated by air gaps to avoid mixing of the samples within the capillary, was optimized and demonstrated.This study shows the potential of ANDeS in laboratory applications by using it for the collection and transfer of biological samples, contained in nanoliter droplets, for subsequent analysis. The experimental results demonstrate the ability of ANDeS to increase the versatility of the DMA platform by allowing for automated retrieval of nanoliter samples from DMA, which was not possible manually on the level of individual droplets. Therefore, it widens the variety of analytical techniques that can be used for the analysis of content of individual droplets and experiments performed using DMA. Thus, ANDeS opens up opportunities to expand the development of miniaturized assays in such fields as cell screening, omics analysis and combinatorial chemistry.

This study examines the intricate relationship between protein glycosylation dynamics and therapeutic responses in Luminal A and Luminal B breast cancer subtypes, focusing on anastrozole and tamoxifen impacts. The present methods inadequately monitor and forecast patient reactions to these treatments, leaving individuals vulnerable to the potential adverse effects of these medications. This research investigated glycan structural changes by following patients for up to 9 months. The protocol involved a series of automated steps including IgG isolation, protein denaturation, glycan labelling, purification, and final analysis using capillary gel electrophoresis with laser-induced fluorescence. The results suggested the significant role of glycan modifications in breast cancer progression, revealing distinctive trends in how anastrozole and tamoxifen elicit varied responses. The findings indicate anastrozole’s association with reduced sialylation and increased core fucosylation, while tamoxifen correlated with increased sialylation and decreased core fucosylation. These observations suggest potential immunomodulatory effects: anastrozole possibly reducing inflammation and tamoxifen impacting immune-mediated cytotoxicity. This study strongly emphasizes the importance of considering specific glycan traits to comprehend the dynamic mechanisms driving breast cancer progression and the effects of targeted therapies. The nuanced differences observed in glycan modifications between these two treatments underscore the necessity for further comprehensive research aimed at thoroughly evaluating the long-term implications and therapeutic efficacy for breast cancer patients.

Lab-on-a-chip technologies and microfluidics have pushed miniaturized liquid handling to unprecedented precision, integration, and automation, which improved the reaction efficiency of immunoassays. However, most microfluidic immunoassay systems still require bulky infrastructures, such as external pressure sources, pneumatic systems, and complex manual tubing and interface connections. Such requirements prevent plug-and-play operation at the point-of-care (POC) settings. Here we present a fully automated handheld general microfluidic liquid handling automation platform with a plug-and-play ‘clamshell-style’ cartridge socket, a miniature electro-pneumatic controller, and injection-moldable plastic cartridges. The system achieved multi-reagent switching, metering, and timing control on the valveless cartridge using electro-pneumatic pressure control. As a demonstration, a SARS-CoV-2 spike antibody sandwich fluorescent immunoassay (FIA) liquid handling was performed on an acrylic cartridge without human intervention after sample introduction. A fluorescence microscope was used to analyze the result. The assay showed a limit of detection at 31.1 ng/mL, comparable to some previously reported enzyme-linked immunosorbent assays (ELISA). In addition to automated liquid handling on the cartridge, the system can operate as a 6-port pressure source for external microfluidic chips. A rechargeable battery with a 12 V 3000 mAh capacity can power the system for 42 h. The footprint of the system is 16.5 × 10.5 × 7 cm, and the weight is 801 g, including the battery. The system can find many other POC and research applications requiring complex liquid manipulation, such as molecular diagnostics, cell analysis, and on-demand biomanufacturing.

Background: Due to supply chain shortages of reagents for real-time (RT)-PCR for SARS-CoV-2 and increasing demand on technical staff, an end-to-end data automation strategy for SARS-CoV-2 sample pooling and singleton analysis became necessary in the summer of 2020. Methods: Using entirely open source software tools—Linux, bash, R, RShiny, ShinyProxy, and Docker—we developed a modular software application stack to manage the preanalytical, analytical, and postanalytical processes for singleton and pooled testing in a 5-week time frame. Results: Pooling was operationalized for 81 days, during which time 64 pooled runs were performed for a total of 5320 sample pools and approximately 21 280 patient samples in 4:1 format. A total of 17 580 negative pooled results were released in bulk. After pooling was discontinued, the application stack was used for singleton analysis and modified to release all viral RT-PCR results from our laboratory. To date, 236 109 samples have been processed avoiding over 610 000 transcriptions. Conclusions: We present an end-to-end data automation strategy connecting 11 devices, one network attached storage, 2 Linux servers, and the laboratory information system. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Over the last decade, whole transcriptome profiling, also known as RNA-sequencing (RNA-seq), has quickly gained traction as a reliable method for unbiased assessment of gene expression. Integration of RNA-seq expression data into other omics datasets (e.g., proteomics, metabolomics, or epigenetics) solidifies our understanding of cell-specific regulatory patterns, yielding pathways to investigate the key rules of gene regulation. A limitation to efficient, at-scale utilization of RNA-seq is the time-demanding library preparation workflows, which is a 2-day or longer endeavor per cohort/sample size.To tackle this bottleneck, we designed an automated workflow that increases throughput capacity, while minimizing human error to enhance reproducibility. To this end, we converted the manual protocol of the NEBNext Directional Ultra II RNA Library Prep Kit for Illumina on the Beckman Coulter liquid handler, Biomek i7 Hybrid workstation. A total of 84 RNA samples were isolated from two human cell lines and subjected to comparative manual and automated library preparation methods.Qualitative and quantitative results indicated a high degree of similarity between libraries generated manually or through automation. Yet, there was a significant reduction in both hands-on and assay time from a 2-day manual to a 9-hour automated workflow. Using linear regression analysis, we found the Pearson correlation coefficient between libraries generated manually or by automation to be almost identical to a sample being sequenced twice (R²= 0.985 vs 0.983). This demonstrates that high-throughput automated workflows can be of great benefit to genomic laboratories by enhancing efficiency of library preparation, reducing hands-on time and increasing throughput potential.

Screening for islet autoantibody markers to identify individuals who are at high risk for developing type 1 diabetes (T1D), often years in advance of clinical symptoms, is both a challenge and a necessity. Identifying high-risk individuals not only reduces hospitalization and rates of life-threatening diabetes ketoacidosis (DKA), but also directs enrollment into prevention trials that require patients who are in the early stages of disease. Here we describe an automated high-throughput multiplex islet autoantibody assay that integrates antibody detection by agglutination-PCR (ADAP) chemistry on the Hamilton Microlab STAR liquid handling platform. The automated system features on-deck thermal cycling and plate sealing to minimize the level of human intervention. The automated multiplex ADAP T1D assay performed similarly to that of manual methods using two distinct cohorts of clinical specimens obtained from the Lucile Packard Children's Hospital at Stanford University and the 2018 Islet Autoantibody Standardization Program (IASP). Notably, the automated assay requires only 4 μL of serum sample for the simultaneous analysis of GAD, IA-2 and insulin autoantibodies.Up to 96 samples may be processed in as little as 3 hours, and the only user intervention required is to transfer a final sealed 96-well plate containing PCR amplicons onto a quantitative PCR (RT-qPCR) instrument for quantification. The automated system is particularly well suited for large-scale analysis of islet autoantibodies in a reproducible, timely, and cost-effective manner.

We report on the fabrication of a syringe-based platform for automation of a colorimetric malaria-Ab assay. We assembled this platform from inexpensive disposable plastic syringes, plastic tubing, easily-obtainable servomotors, and an Arduino microcontroller chip, which allowed for system automation. The automated system can also be fabricated using stereolithography (SLA) to print elastomeric reservoirs (used instead of syringes), while platform framework, including rack and gears, can be printed with fused deposition modeling (FDM). We report on the optimization of FDM and SLA print parameters, as well as post-production processes. A malaria-Ab colorimetric test was successfully run on the automated platform, with most of the assay reagents dispensed from syringes. Wash solution was dispensed from an SLA-printed elastomeric reservoir to demonstrate the feasibility of both syringe and elastomeric reservoir-based approaches. We tested the platform using a commercially available malaria-Ab colorimetric assay originally designed for spectroscopic plate readers. Unaided visual inspection of the assay solution color change was sufficient for qualitative detection of positive and negative samples. A smart phone application can also be used for quantitative measurement of the assay color change.

Polymerase chain reaction (PCR)-based assays were widely deployed during the SARS-CoV-2 pandemic for population-scale testing. High-throughput molecular diagnostic laboratories required a high degree of process automation to cope with huge testing demands, fast turnaround times, and quality requirements. However, process developers and optimizers often neglected the critical step of preparing a PCR Master Mix. The construction of PCR Master Mix depends on operator skill during the manual pipetting of reagents. Manual procedures introduce variation, inconsistency, wastage, and potentially risks data integrity. To address this, we developed a liquid-handler-based solution for automated, traceable, and compliant PCR Master Mix preparation. Here, we show that a fully automated PCR Master Mix protocol can replace manual pipetting, even in a diagnostic environment, without affecting accuracy or precision. Ultimately, this method eliminated operator-induced wastage and improved the consistency of the quality of results., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Giorgio Fedele reports equipment, drugs, or supplies was provided by SPT Labtech Ltd. Giorgio Fedele reports equipment, drugs, or supplies was provided by LGC Biosearch Technologies Inc. Giorgio Fedele reports equipment, drugs, or supplies was provided by BMG LABTECH GmbH. Donald Fraser reports financial support was provided by United Kingdom Department of Health and Social Care. Denise Grovewood reports a relationship with SPT Labtech Ltd that includes: employment. The authors have no further conflicts of interest to declare. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Fluorescent in situ hybridization (FISH) can provide spatial information about DNA/RNA targets in fixed cells and tissues. However, the workflows of multiplexed FISH-based imaging that use sequential rounds of hybridization quickly become laborious as the number of rounds increases because of liquid handling demands. Here, we present an open-source and low-cost fluidics system that is purpose built for automating the workflows of sequential FISH-based imaging. Our system features a fluidics module with 16 addressable channels in which flow is positive pressure-driven and switched on/off by solenoid valves in order to transfer FISH reagents to the sample. Our system also includes a controller with a main printed circuit board that can control up to 120 solenoid valves and allows users to control the fluidics module via serial communication. We demonstrate the automatic and robust fluid exchange with this system by targeting the alpha satellite repeat in HeLa cell with 14 rounds of sequential hybridization and imaging. We anticipate that this simple and flexible system will be of utility to researchers performing multiplexed in situ assays in a range of experimental systems.

Droplet-based microfluidic screening techniques can benefit from interfacing established microtiter plate-based screening and sample management workflows. Interfacing tools are required both for loading preconfigured microtiter-plate (MTP)-based sample collections into droplets and for dispensing the used droplets samples back into MTPs for subsequent storage or further processing. Here, we present a collection of Digital Microfluidic Pipetting Tips (DMPTs) with integrated facilities for droplet generation and manipulation together with a robotic system for its operation. This combination serves as a bidirectional sampling interface for sample transfer from wells into droplets (w2d) and vice versa droplets into wells (d2w). The DMPT were designed to fit into 96-deep-well MTPs and prepared from glass by means of microsystems technology. The aspirated samples are converted into the channel-confined droplets' sequences separated by an immiscible carrier medium. To comply with the demands of dose-response assays, up to three additional assay compound solutions can be added to the sample droplets. To enable different procedural assay protocols, four different DMPT variants were made. In this way, droplet series with gradually changing composition can be generated for, e.g., 2D screening purposes. The developed DMPT and their common fluidic connector are described here. To handle the opposite transfer d2w, a robotic transfer system was set up and is described briefly. [ABSTRACT FROM AUTHOR]

Using established needle‐ and tip‐based liquid‐handlers partially restricts applications that require the handling of greater sample amounts, options for filtration or handling of liquids with uncommon behavior. In this paper the prototype development of a versatile liquid pouring device (vLiPD) is described, which offers complementary features to usual liquid‐handling solutions. For mapping the container characteristics, a geometrical and an on 3D‐CAD‐model‐based approach are introduced and compared. Influences by the liquids' behaviors are compensated by liquid‐specific calibration factors. [ABSTRACT FROM AUTHOR]

In the life sciences, automation solutions are primarily established in the field of drug discovery. However, there is also an increasing need for automated solutions in the field of medical diagnostics, e.g., for the determination of vitamins, medication or drug abuse. While the actual metrological determination is highly automated today, the necessary sample preparation processes are still mainly carried out manually. In the laboratory, flexible solutions are required that can be used to determine different target substances in different matrices. A suitable system based on an automated liquid handler was implemented. It has been tested and validated for the determination of three cannabinoid metabolites in blood, urine and saliva. To extract Δ9-tetrahydrocannabinol-D3 (Δ9-THC-D3), 11-hydroxy-Δ9-tetrahydrocannabinol (THC-OH) and 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THC-COOH) from serum, urine and saliva both rapidly and cost-effectively, three sample preparation methods automated with a liquid handling robot are presented in this article, the basic framework of which is an identical SPE method so that they can be quickly exchanged against each other when the matrix is changed. If necessary, the three matrices could also be prepared in parallel. For the sensitive detection of analytes, protein precipitation is used when preparing serum before SPE and basic hydrolysis is used for urine to cleave the glucuronide conjugate. Recoveries of developed methods are >77%. Coefficients of variation are <4%. LODs are below 1 ng/mL and a comparison with the manual process shows a significant cost reduction. [ABSTRACT FROM AUTHOR]

The implementation of automation has already had a considerable impact on chemical and pharmaceutical industrial laboratories. However, academic laboratories have often been more reluctant to adopt such technology due to the high cost of commercial liquid handling systems, although, in many instances, there would be a huge potential to automate repetitive tasks, resulting in elevated productivity. We present here a detailed description of the setup, validation, and utilization of a multifunctional liquid automation (MULA) system that can be used to automate various chemical and biological tasks. Considering that such a setup must be highly customizable, we also designed MULA with respect to modularity, providing detailed insight as far as possible. Including all 3D-printed parts and the used Hamilton gastight micro syringe, the total construction cost is approximately 700 €. This allows us to achieve a highly reliable and accurate system that exceeds the precision of a classical air displacement pipette while still retaining the ability to use closed vial (septa) setups. To encourage other groups to adopt this setup, detailed instructions and tips for every step of the process are provided, along with the complete CAD design of MULA and control code, which are freely available for download under the CC BY NC 3.0 license., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Author(s).)

Many routines in biological experiments require the precise handling of liquid volumes in the range of microliters up to liters. In this paper, we describe a new wireless controller that is adapted to liquid manipulation tasks, in particular when combined with the proposed 3D-printed pumps. It can be built from widely available electronic components and managed with open-source software. The use of peristaltic pumps enables to move volumes from milliliters to liters with a relative error below 1% or a syringe pump capable of injecting volumes in the range of milliliters with microliter accuracy. The system is remotely controllable over WiFi and easily automated using the MQTT communication protocol. The programming of the microcontroller is performed on the Arduino IDE. The WiFi settings and the calibration value can be easily modified, stored and exported in the form of a JSON file to create a user friendly, plug and play and easily scalable device. Additional sensors or actuators can be added, allowing the system to adapt to various usages. Finally, in addition to its low manufacturing cost and its capability to fit a large variety of tasks involving liquid handling, our system has been specifically designed for research environments where adaptability and repeatability of experiments is essential.

Every year biotechnology labs generate a combined total of ∼5.5 million tons of plastic waste. As the global bioeconomy expands, biofoundries will inevitably increase plastic consumption in-step with synthetic biology scaling. Decontamination and reuse of single-use plastics could increase sustainability and reduce recurring costs of biological research. However, throughput and variable cleaning quality make manual decontamination impractical in most instances. Automating single-use plastic cleaning with liquid handling robots makes decontamination more practical by offering higher throughput and consistent cleaning quality. However, open-source, validated protocols using low-cost lab robotics for effective decontamination of plasticware-facilitating safe reuse-have not yet been developed. Here we introduce and validate TidyTron: a library of protocols for cleaning micropipette tips and microtiter plates that are contaminated with DNA, E. coli, and S. cerevisiae. We tested a variety of cleaning solutions, contact times, and agitation methods with the aim of minimizing time and cost, while maximizing cleaning stringency and sustainability. We tested and validated these cleaning procedures by comparing fresh (first-time usage) versus cleaned tips and plates for contamination with cells, DNA, or cleaning solutions. We assessed contamination by measuring colony forming units by plating, PCR efficiency and DNA concentration by qPCR, and event counts and debris by flow cytometry. Open source cleaning protocols are available at https://github.com/PlantSynBioLab/TidyTron and hosted on a graphical user interface at https://jbryantvt.github.io/TidyTron/., Competing Interests: Declaration of Competing Interest The authors declare no conflicts of interest., (Copyright © 2023. Published by Elsevier Inc.)

Automated water sampling can be very useful, but open-source choices are limited. Here I present an autosampler which consists of a gantry robot that delivers water from a syringe pump to 24 capped 40 ml vials. The autosampler is controlled using an Android tablet automatized using Macrodroid. Three rinsing cycles ensure negligible carryover between consecutive samples. Hourly sampling from a creek under rainy conditions suggested that total organic carbon in water was diluted by the rain. Some important limitations: 1) the autosampler must be on a steady, flat, horizontal surface; 2) unattended sampling can only last as long as the batteries powering the tablet and the motors; 3) distance from the syringe pump to water cannot exceed ~2 m in height and ~4 m in length for 3 mm tubing; 4) sampling frequency does not exceed one sample every eleven minutes. However, because of its open design, the autosampler can be modified and improved to not only overcome these limitations, but also potentially expand its scope to more demanding sampling if necessary.

In the life sciences, automation solutions are primarily established in the field of drug discovery. However, there is also an increasing need for automated solutions in the field of medical diagnostics, e.g., for the determination of vitamins, medication or drug abuse. While the actual metrological determination is highly automated today, the necessary sample preparation processes are still mainly carried out manually. In the laboratory, flexible solutions are required that can be used to determine different target substances in different matrices. A suitable system based on an automated liquid handler was implemented. It has been tested and validated for the determination of three cannabinoid metabolites in blood, urine and saliva. To extract Δ9-tetrahydrocannabinol-D3 (Δ9-THC-D3), 11-hydroxy-Δ9-tetrahydrocannabinol (THC-OH) and 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THC-COOH) from serum, urine and saliva both rapidly and cost-effectively, three sample preparation methods automated with a liquid handling robot are presented in this article, the basic framework of which is an identical SPE method so that they can be quickly exchanged against each other when the matrix is changed. If necessary, the three matrices could also be prepared in parallel. For the sensitive detection of analytes, protein precipitation is used when preparing serum before SPE and basic hydrolysis is used for urine to cleave the glucuronide conjugate. Recoveries of developed methods are >77%. Coefficients of variation are

Chitosan has various merits including antimicrobial activity but shows poor liquid handling properties such as fluid absorption and retention, keeping moisture environment, and gel-formation. In order to improve the characteristics of chitosan nonwovens as a wound dressing material, carboxymethyl chitosan nonwovens were prepared by direct carboxymethylation reaction of needle-punched chitosan nonwovens. FT-IR and degree of deacetylation analysis showed O-substitution was dominant, and the amount of sodium chloroacetate and sodium hydroxide had effects on degree of substitution. As the degree of substitution increased, fluid absorption capacities, fluid retention capacities, wet dimensional stability, and tensile strength were increased. Liquid spreading and moisture evaporation were suppressed with the increase of degree of substitution. Overall, O-carboxymethyl chitosan has good potential as a moist wound dressing material. [ABSTRACT FROM AUTHOR]

The aim of this study was to semi-automate process analytics for the quantification of common impurities in downstream processing such as host cell DNA, host cell proteins and endotoxins using a commercial liquid handling station. By semi-automation, the work load to fully analyze the elution peak of a purification run was reduced by at least 2.41 h. The relative standard deviation of results among different operators over a time span of up to 6 months was at the best reduced by half, e.g. from 13.7 to 7.1% in dsDNA analysis. Automation did not improve the reproducibility of results produced by one operator but released time for data evaluation and interpretation or planning of experiments. Overall, semi-automation of process analytics reduced operator-specific influence on test results. Such robust and reproducible analytics is fundamental to establish process analytical technology and get downstream processing ready for Quality by Design approaches. [ABSTRACT FROM AUTHOR]

The increasing emphasis on portability and miniaturization in biomedical analysis and detection necessitates higher accuracy and applicability in handling sample reagents. In this study, we propose a high-precision portable programmable nanopipette based on pneumatic drop-on-demand and gas displacement pipetting techniques. The nanopipette employs programmable electropneumatic regulators and solenoid valves for precise control over air pressure, air pulse duration, and the number of air pulses, enabling accurate dispensing of specific liquid volumes. The nanopipette provides both discrete and continuous dispensing modes. In the discrete mode, it employs a single droplet as the minimum unit, allowing for precise dispensing of sub-microliter liquids (≤500 nL) by accumulating uniformly sized droplets. Conversely, the continuous mode enables the rapid dispensing of larger volume liquids (>500 nL) through continuous gas displacement. Thanks to the two dispensing modes, the nanopipette can handle a broad range of dispensing volumes, spanning six orders of magnitude (from 3 nL to 1 mL), with a minimum volume increment of 1 nL. Through validation using two characterization methods, the nanopipette demonstrates systematic errors (random errors) ranging from − 0.5% to − 3.6% (± 0.9% to ± 4.4%), surpassing the performance of existing commercial micropipettes. In proof-of-concept experiments involving extensive dilution and quantitative analysis of drug toxicity, the nanopipette efficiently simplified experimental procedures (from multiple dilutions to single-step dilution), minimized reagent consumption (from 22 mL to 500 μL), and produced highly accurate and reliable experimental results. These findings highlight the applicability and potential value of the nanopipette in the field of biomedical research. [Display omitted] • A high-precision, wide volume range nanopipette based on drop-on-demand technique and gas displacement pipetting technique. • Nanopipette enables liquid handling within the range of 3 nL to 1 mL, with a minimum volume increment of 1 nL. • The biocompatibility and applicability of nanopipette have been demonstrated in biomedical experiments. [ABSTRACT FROM AUTHOR]

The Droplet Microarray (DMA) has emerged as a tool for high-throughput biological and chemical applications by enabling miniaturization and parallelization of experimental processes. Due to its ability to hold hundreds of nanoliter droplets, the DMA enables simple screening and analysis of samples such as cells and biomolecules. However, handling of nanoliter volumes poses a challenge, as manual recovery of nanoliter volumes is not feasible, and traditional laboratory equipment is not suited to work with such low volumes, and small array formats. To tackle this challenge, we developed the Automated Nanoliter Droplet Selection device (ANDeS), a robotic system for automated collection and transfer of nanoliter samples from DMA. ANDeS can automatically collect volumes from 50 to 350 nL from the flat surface of DMA with a movement accuracy of ±30 µm using fused silica capillaries. The system can automatically collect and transfer the droplets from DMA chip into other platforms, such as microtiter plates, conical tubes or another DMA. In addition, to ensure high throughput and multiple droplet collection, the uptake of multiple droplets within a single capillary, separated by air gaps to avoid mixing of the samples within the capillary, was optimized and demonstrated. This study shows the potential of ANDeS in laboratory applications by using it for the collection and transfer of biological samples, contained in nanoliter droplets, for subsequent analysis. The experimental results demonstrate the ability of ANDeS to increase the versatility of the DMA platform by allowing for automated retrieval of nanoliter samples from DMA, which was not possible manually on the level of individual droplets. Therefore, it widens the variety of analytical techniques that can be used for the analysis of content of individual droplets and experiments performed using DMA. Thus, ANDeS opens up opportunities to expand the development of miniaturized assays in such fields as cell screening, omics analysis and combinatorial chemistry., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Joaquin E. Urrutia Gomez reports financial support was provided by German Academic Exchange Service. Anna A. Popova reports financial support was provided by Impuls und Vernetzungsfonds der Helmholtz-Gemeinschaft, Heidelberg and Karlsruhe cooperation program. Pavel A. Levkin reports financial support was provided by German Research Foundation. Pavel A. Levkin reports a relationship with Aquarray GmbH that includes: board membership. Anna A. Popova reports a relationship with Aquarray GmbH that includes: board membership., (Copyright © 2023. Published by Elsevier Inc.)

Composite nonwovens were prepared by the lamination of polypropylene spunbond nonwoven with carboxymethyl cellulose/hollow viscose rayon blend nonwoven and their various properties, such as liquid handling properties, wet tensile strength, moisture evaporation rate, and in-vitro cytotoxicity, were evaluated for moist wound dressing applications. Fluid retention capacity, wet dimensional stability, and wet tensile strength of carboxymethyl cellulose/hollow viscose rayon nonwoven were significantly improved by the lamination with polypropylene spunbond nonwoven. The polypropylene spunbond nonwoven layer did not affect the liquid spreadability and moisture evaporation. The composite nonwoven was not cytotoxic. Considering liquid handling properties and wet properties, composite nonwoven, in which polypropylene spunbond nonwoven on both sides of carboxymehtyl cellulose/hollow viscose rayon nonwoven, was found to be the most desirable wound dressing material of those assessed in this study. [ABSTRACT FROM AUTHOR]

Abstract The ability to engineer monoclonal antibodies (mAbs) with high specificity made mAbs the fastest growing segment in the drug market. mAbs represent 8 of the top 20 selling drugs with combined sales of more than 57 billion US$ per year. The ability to purify large numbers of mAbs with sufficient yields for initial screening campaigns has direct impact on the timelines of a project. Automated liquid handling (ALH)-based mAb purification platforms have been used to facilitate the production of large numbers of mAbs. However, the ongoing pressure to de-risk potential lead molecules at an early development stage by including bio-physical characterization of mAbs has further increased the demand to produce sufficient quantities from limited sample volumes. A bottleneck so far has been the limited dynamic binding capacity of these systems, which is partly due to the binding properties of commonly used Protein A affinity matrices. The present publication suggests that by using a Protein A matrix optimized for continuous chromatography applications the yields of ALH-based but also standard lab-scale mAb purifications can be significantly increased without the need to change established protocols. Highlights • Applied continuous chromatography protein A matrix to robotic mAb purification. • Dynamic binding capacity increased from 15 mg/ml to more than 40 mg/ml. • Preparative robotic mAb purification with increased yields for subsequent characterization. • Significantly increased mAb yields on lab scale instruments. [ABSTRACT FROM AUTHOR]

The fruit fly, 'Drosophila melanogaster', continues to be one of the most widely used model organisms in biomedical research. Though chosen for its ease of husbandry, maintaining large numbers of stocks of fruit flies, as done by many laboratories, is labour-intensive. One task which lends itself to automation is the production of the vials of food in which the flies are reared. Fly facilities typically have to generate several thousand vials of fly food each week to sustain their fly stocks. The system presented here combines a cartesian coordinate robot with a peristaltic pump. The design of the robot is based on an open hardware CNC (computer numerical control) machine, and uses belt and pulley actuators for the X and Y axes, and a leadscrew actuator for the Z axis. CNC motion and operation of the peristaltic pump are controlled by grbl (gnea 2018), an open source, embedded, G-code parser. Grbl is written in optimized C and runs directly on an Arduino. A Raspberry Pi is used to generate and stream G-code instructions to Grbl. A touch screen on the Raspberry Pi provides a graphical user interface to the system. Whilst the robot was built for the express purpose of filling vials of fly food, it could potentially be used for other liquid handling tasks in the laboratory.

Programmable liquid handling devices for cell culture systems have dramatically enhanced scalability and reproducibility. We previously reported a protocol to produce cell aggregates demonstrating growth plate-like structures containing hypertrophic chondrocytes from human induced pluripotent stem cells (hiPSCs). To apply this protocol to large-scale drug screening for growth plate-related diseases, we adapted it to the automated cell culture system (ACCS) consisting of programmable liquid handling devices connected to CO 2 incubators, a refrigerator, and labware feeders, designed for up to 4 batches with several cell culture plates culturing for several months. We developed a new program preparing culture media with growth factors at final concentration immediately before dispensing them to each well and precisely positioning the tip for the medium change without damaging cell aggregates. Using these programs on the ACCS, we successfully cultured cell aggregates for 56 days, only needing to replenish the labware, medium, and growth factors twice a week. The size of cell aggregates in each well increased over time, with low well-to-well variability. Cell aggregates on day 56 showed histochemical, immunohistochemical, and gene expression properties of growth plate-like structures containing hypertrophic chondrocytes, indicating proper quality as materials for basic research and drug discovery of growth plate related diseases. The established program will be a suitable reference for making programs of experiments requiring long term and complex culture procedures using ACCS., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Inc.)

Consistent and accurate manipulation of liquids is essential for many biology experiments. Teaching these skills is challenging in the laboratory. I have developed Training Tips, a homemade teaching tool to train students to use micropipettes in the classroom and laboratory. Training Tips provide obvious visual targets that enable inexperienced students to know immediately if they are pipetting accurately. [ABSTRACT FROM AUTHOR]

We report on the fabrication of a syringe-based platform for automation of a colorimetric malaria-Ab assay. We assembled this platform from inexpensive disposable plastic syringes, plastic tubing, easily-obtainable servomotors, and an Arduino microcontroller chip, which allowed for system automation. The automated system can also be fabricated using stereolithography (SLA) to print elastomeric reservoirs (used instead of syringes), while platform framework, including rack and gears, can be printed with fused deposition modeling (FDM). We report on the optimization of FDM and SLA print parameters, as well as post-production processes. A malaria-Ab colorimetric test was successfully run on the automated platform, with most of the assay reagents dispensed from syringes. Wash solution was dispensed from an SLA-printed elastomeric reservoir to demonstrate the feasibility of both syringe and elastomeric reservoir-based approaches. We tested the platform using a commercially available malaria-Ab colorimetric assay originally designed for spectroscopic plate readers. Unaided visual inspection of the assay solution color change was sufficient for qualitative detection of positive and negative samples. A smart phone application can also be used for quantitative measurement of the assay color change.

Lab-on-a-chip technologies and microfluidics have pushed miniaturized liquid handling to unprecedented precision, integration, and automation, which improved the reaction efficiency of immunoassays. However, most microfluidic immunoassay systems still require bulky infrastructures, such as external pressure sources, pneumatic systems, and complex manual tubing and interface connections. Such requirements prevent plug-and-play operation at the point-of-care (POC) settings. Here we present a fully automated handheld general microfluidic liquid handling automation platform with a plug-and-play 'clamshell-style' cartridge socket, a miniature electro-pneumatic controller, and injection-moldable plastic cartridges. The system achieved multi-reagent switching, metering, and timing control on the valveless cartridge using electro-pneumatic pressure control. As a demonstration, a SARS-CoV-2 spike antibody sandwich fluorescent immunoassay (FIA) liquid handling was performed on an acrylic cartridge without human intervention after sample introduction. A fluorescence microscope was used to analyze the result. The assay showed a limit of detection at 31.1 ng/mL, comparable to some previously reported enzyme-linked immunosorbent assays (ELISA). In addition to automated liquid handling on the cartridge, the system can operate as a 6-port pressure source for external microfluidic chips. A rechargeable battery with a 12 V 3000 mAh capacity can power the system for 42 h. The footprint of the system is 16.5 × 10.5 × 7 cm, and the weight is 801 g, including the battery. The system can find many other POC and research applications requiring complex liquid manipulation, such as molecular diagnostics, cell analysis, and on-demand biomanufacturing., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Author(s).)

Rapid development of chromatographic processes relies on effective high-throughput screening (HTS) methods. This article describes the development of pseudo-linear gradient elution for resin selectivity screening using RoboColumns®. It gives guidelines for the implementation of this HTS method on a Tecan Freedom EVO® robotic platform, addressing fundamental aspects of scale down and liquid handling. The creation of a flexible script for buffer preparation and column operation plus efficient data processing provided the basis for this work. Based on the concept of discretization, linear gradient elution was transformed into multistep gradients. The impact of column size, flow rate, multistep gradient design, and fractionation scheme on separation efficiency was systematically investigated, using a ternary model protein mixture. We identified key parameters and defined optimal settings for effective column performance. For proof of concept, we examined the selectivity of several cation exchange resins using various buffer conditions. The final protocol enabled a clear differentiation of resin selectivity on miniature chromatography column (MCC) scale. Distinct differences in separation behavior of individual resins and the influence of buffer conditions could be demonstrated. Results obtained with the robotic platform were representative and consistent with data generated on a conventional chromatography system. A study on antibody monomer/high molecular weight separation comparing MCC and lab scale under higher loading conditions provided evidence of the applicability of the miniaturized approach to practically relevant feedstocks with challenging separation tasks as well as of the predictive quality for larger scale. A comparison of varying degrees of robotic method complexity with corresponding effort (analysis time and labware consumption) and output quality highlights tradeoffs to select a method appropriate for a given separation challenge or analytical constraints. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1503-1519, 2016 [ABSTRACT FROM AUTHOR]

Monoclonal antibodies (mAbs) have become the fastest growing segment in the drug market with annual sales of more than 40 billion US$ in 2013. The selection of lead candidate molecules involves the generation of large repertoires of antibodies from which to choose a final therapeutic candidate. Improvements in the ability to rapidly produce and purify many antibodies in sufficient quantities reduces the lead time for selection which ultimately impacts on the speed with which an antibody may transition through the research stage and into product development. Miniaturization and automation of chromatography using micro columns (RoboColumns ® from Atoll GmbH) coupled to an automated liquid handling instrument (ALH; Freedom EVO ® from Tecan) has been a successful approach to establish high throughput process development platforms. Recent advances in transient gene expression (TGE) using the high-titre Expi293F™ system have enabled recombinant mAb titres of greater than 500 mg/L. These relatively high protein titres reduce the volume required to generate several milligrams of individual antibodies for initial biochemical and biological downstream assays, making TGE in the Expi293F™ system ideally suited to high throughput chromatography on an ALH. The present publication describes a novel platform for purifying Expi293F™-expressed recombinant mAbs directly from cell-free culture supernatant on a Perkin Elmer JANUS-VariSpan ALH equipped with a plate shuttle device. The purification platform allows automated 2-step purification (Protein A—desalting/size exclusion chromatography) of several hundred mAbs per week. The new robotic method can purify mAbs with high recovery (>90%) at sub-milligram level with yields of up to 2 mg from 4 mL of cell-free culture supernatant. [ABSTRACT FROM AUTHOR]