Journal of Laboratory Automation

The clinical team attending to a patient upon a diagnosis is faced with two main questions: what treatment, and at what dose? Clinical trials’ results provide the basis for guidance and support for official protocols that clinicians use to base their decisions upon. However, individuals rarely demonstrate the reported response from relevant clinical trials, often the average from a group representing a population or subpopulation. The decision complexity increases with combination treatments where drugs administered together can interact with each other, which is often the case. Additionally, the individual’s response to the treatment varies over time with the changes in his or her condition, whether via the indication or physiology. In practice, the drug and the dose selection depend greatly on the medical protocol of the healthcare provider and the medical team’s experience. As such, the results are inherently varied and often suboptimal. Big data approaches have emerged as an excellent decision-making support tool, but their application is limited by multiple challenges, the main one being the availability of sufficiently big datasets with good quality, representative information. An alternative approach—phenotypic personalized medicine (PPM)—finds an appropriate drug combination (quadratic phenotypic optimization platform [QPOP]) and an appropriate dosing strategy over time (CURATE.AI) based on small data collected exclusively from the treated individual. PPM-based approaches have demonstrated superior results over the current standard of care. The side effects are limited while the desired output is maximized, which directly translates into improving the length and quality of individuals’ lives.

Recording the data necessary to assess the kinetics of a reaction can be labor-intensive. In this technology brief, we show a method to automate this task by utilizing parts of an ÄKTApurifier chromatography system to automatically take samples from a reaction vessel at predefined time intervals and place them in 96-well plates and also enable correlating the samples with in-line spectral data of the reaction solution. Automatic sampling can reduce experimental bottlenecks by enabling overnight reactions or a higher degree of parallelization. To demonstrate the feasibility of the method, we performed batch-PEGylation of lysozyme with varying conditions by changing the molar excess of the PEG reagent. We used analytical cation-exchange chromatography to analyze the samples taken during the batch reaction, determining the concentrations of the individual species present at each time step. Subsequently, we fitted a kinetic model on these data. Fitting the model to four different reaction conditions simultaneously yielded a regression coefficient of R² = 0.871.

The future of the life sciences is linked to automation and microfluidics. As robots start working side by side with scientists, robotic automation of microfluidics in general, and droplet microfluidics in particular, will significantly extend and accelerate the life sciences. Here, we demonstrate the automation of droplet microfluidics using an inexpensive liquid-handling robot to produce human scaffold-free cell spheroids at high throughput. We use pipette actuation and interface the pipetting tip with a droplet-generating microfluidic device. In this device, we produce highly monodisperse droplets with a diameter coefficient of variation (CV) lower than 2%. By encapsulating cells in these droplets, we produce cell spheroids in droplets and recover them to standard labware containers at a throughput of 85,000 spheroids per microfluidic circuit per hour. The viability of the cells in spheroids remains high throughout the process and decreases by >10% (depending on the cell line used) after a 16 h incubation period in nanoliter droplets and automated recovery. Scaffold-free cell spheroids and 3D tissue constructs recapitulate many aspects of functional human tissue more accurately than 2D or single-cell cultures, but assembly methods for spheroids (e.g., hanging drop microplates) have limited throughput. The increased throughput and decreased cost of our method enable spheroid production at the scale needed for lead discovery drug screening, and approach the cost at which these microtissues could be used as building blocks for organ-scale regenerative medicine.

This work describes the use of electrothermal atomic absorption spectrometry in combination with a pyrolytic graphite-coated tube with a platform for trace arsenic (As) determination in titanium dioxide (TiO₂) pigment. This type of matrix is challenging, as complete digestion in hydrofluoric acid-containing solution is needed. First, closed-vessel microwave digestion was performed for the full-sample decomposition. Next, a temperature program was optimized for drying, pyrolysis, and atomization temperatures. Furthermore, the use of a chemical modifier mixture was proposed that reduced the background contribution and prevented significant analyte loss and therefore improved the analytical procedure. The optimized method was validated for the detection (LOD) and quantification (LOQ) limits, the linear concentration range, accuracy, and precision. Special attention was devoted to the matrix-matching solutions in the calibration procedure. Linearity was confirmed in the 5.0 to 100.0 µg/L concentration range (R² = 0.999). The average recovery for 16 different real TiO₂ pigment samples was 92.0%, and the relative standard deviation value for six replicate measurements was ≤10.4%. Moreover, the LOD and LOQ in terms of the TiO₂ pigment mass was determined to be 0.2 µg/(g TiO₂) and 0.7 µg/(g TiO₂), respectively. The latter complies with Commission Directive 2008/128/EC, which does not allow more than 3 µg As/(g product) as the specific criteria of purity. Finally, based on scanning electron microscopy analysis of unused and several times used pyrolytic graphite-coated tubes, usage of the tube 250 times before replacement is recommended.

This paper presents a droplet-based immunoassay chip allowing each droplet to be positioned in a passive droplet-positioning cavern under continuous flow. In addition, the chip surface can immobilize any kind of histidine-tagged capture agents for performing simultaneous multiplex immunoassays. Distinct families of monodispersed droplets were generated since a diaphragm, which is a thin elastomeric flap film suspended from the top of the main channel, forms a double T junction for shearing the aqueous liquids by the carrier flow. These two types of monodispersed droplets traverse the main channel to the downstream detection area and enter the passive positioning caverns for further immunoassay. A layer of Ni–Co film was coated on the substrate by electrodeposition in order to immobilize the multiplex histidine-tagged capture molecules. In this study, the tumor suppressor protein p53 and the extracellular signal-related kinase 1 (ERK1) were used as the capture agents. Then, both histidine-tagged proteins p53 and ERK1 were immobilized by the Ni–Co layer in a microarray format for subsequent immunoassay and fluorescence detection. The experimental results show that the detected fluorescence intensity is proportioned to the concentration of the encapsulated content in a small droplet. This proposed droplet-based immunoassay chip can immobilize multiplex histidine-tagged proteins, irrelevant to the species of proteins, to carry out simultaneous immunoassays and allow the operation sequence to be conducted automatically through the manipulation of droplets.

Droplet-based microfluidics holds enormous potential for transforming high-throughput drug screening. Miniaturization through droplets in combination with automation contributes to reduce reagent use and analysis time as well as minimizing or eliminating labor-intensive steps leading to associated reductions in cost. In this paper, we demonstrate the potential of automated and cost-effective microfluidic droplet-generating technology in the context of an enzymatic activity assay for screening collagenase inhibitors. Experimental results show reproducible and accurate creation and mixing of droplet combinations resulting in biochemical data comparable to data produced by an industry standard instrument. This microfluidic platform that can generate and combine multiple droplets represents a promising tool for high-throughput drug screening.

The emphasis of this paper lies in the fabrication of a three-layer polydimethylsiloxane chip for micro liquid sample operation. In this paper, the microchannels with a rectangular control layer cross section are fabricated based on a dry-film negative photoresist mold, while the microchannels with a rounded liquid layer cross section are fabricated by a positive photoresist reflow mold. The relationships between temperature and the time of reflow and the arc level of the liquid layer mold are discussed. Different ratios, curing temperatures, and curing times are used to fabricate the two PDMS layers to improve their toughness and plasticity separately. The PDMS slabs with microstructure networks are treated with oxygen plasma to improve their surface properties. The improved surface properties serve to reduce the temperature and time, and improve the sealing strength, which is as effective as adding PDMS in varying ratios. The micro liquid sample operation experiments show that high levels of pinching off and mixing performances on pneumatic microfluidic chips are obtained more easily.

Comprehensive analysis of tumor heterogeneity requires robust methods for the isolation and analysis of single cells from patient samples. An ideal approach would be fully compatible with downstream analytic methods, such as advanced genomic testing. These endpoints necessitate the use of live cells at high purity. A multitude of microfluidic circulating tumor cell (CTC) enrichment technologies exist, but many of those perform bulk sample enrichment and are not, on their own, capable of single-cell interrogation. To address this, we developed an affordable semiautomated single-cell aspirator (SASCA) to further enrich rare-cell populations from a specialized microwell array, per their phenotypic markers. Immobilization of cells within microwells, integrated with a real-time image processing software, facilitates the detection and precise isolation of targeted cells that have been optimally seeded into the microwells. Here, we demonstrate the platform capabilities through the aspiration of target cells from an impure background population, where we obtain purity levels of 90%–100% and demonstrate the enrichment of the target population with high-quality RNA extraction. A range of low cell numbers were aspirated using SASCA before undergoing whole transcriptome and genome analysis, exhibiting the ability to obtain endpoints from low-template inputs. Lastly, CTCs from patients with castration-resistant prostate cancer were isolated with this platform and the utility of this method was confirmed for rare-cell isolation. SASCA satisfies a need for an affordable option to isolate single cells or highly purified subpopulations of cells to probe complex mechanisms driving disease progression and resistance in patients with cancer.

The primary goal of bioprocess cell line development is to obtain high product yields from robustly growing and well-defined clonal cell lines in timelines measured in weeks rather than months. Likewise, high-throughput screening of B cells and hybridomas is required for most cell line engineering workflows. A substantial bottleneck in these processes is detecting and isolating rare clonal cells with the required characteristics. Traditionally, this was achieved by the resource-intensive method of limiting dilution cloning, and more recently aided by semiautomated technologies such as cell sorting (e.g., fluorescence-activated cell sorting) and colony picking. In this paper we report on our novel Cyto-Mine Single Cell Analysis and Monoclonality Assurance System, which overcomes the limitations of current technologies by screening hundreds of thousands of individual cells for secreted target proteins, and then isolating and dispensing the highest producers into microtiter plate wells (MTP). The Cyto-Mine system performs this workflow using a fully integrated, microfluidic Cyto-Cartridge. Critically, all reagents and Cyto-Cartridges used are animal component-free (ACF) and sterile, thus allowing fast, robust, and safe isolation of desired cells.

3D-printed laboratory devices can enable ambitious research purposes even at a low-budget level. To follow this trend, here we describe the construction, calibration, and usage of the FINDUS (Fully Integrable Noncommercial Dispensing Utility System). We report the successful 3D printing and assembly of a liquid-handling workstation for less than $400. Using this setup, we achieve reliable and flexible liquid-dispensing automation with relative pipetting errors of less than 0.3%. We show our system is well suited for several showcase applications from both the biology and chemistry fields. In support of the open-source spirit, we make all 3D models, assembly instructions, and source code available for free download, rebuild, and modification.

The demand for efficient and qualified measurements is high. The measurements of physical parameters in industrial environments usually are automatically executed using process measurement technologies. The close coupling of the measurement system and process equipment enables in-process data acquisition (inline or online). In contrast, compound-oriented measurements for the qualitative identification and quantitative determination of chemical elements and chemical compounds, or the determination of biological behavior, are still challenging in measurement processes. Today, biological assays as well as high-content and high-throughput screening procedures are well automated. It appears quite differently if the efficiency of analytical measurements should be improved. Due to the characteristics of the samples and processes, inline or online coupling is often not possible. Therefore, atline or offline couplings are gaining in importance. In combination with robotic systems, efficient automation of analytical measurements can be reached.

The book Automation Solutions for Analytical Measurements: Concepts and Applications presents and discusses suitable automation concepts and a variety of realized robot-based analytical measurement systems. In this commentary, the main findings are highlighted and discussed.