HardwareX
○ Elsevier BV
Preprints posted in the last 90 days, ranked by how well they match HardwareX's content profile, based on 18 papers previously published here. The average preprint has a 0.02% match score for this journal, so anything above that is already an above-average fit.
Versluis, D. M.; Benninger, M.; Panigrahi, D. P.; Hoque, A.; Machesky, L. M.; Tiengwe, C.; Insall, R.
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Petri dishes are routinely photographed using handheld cameras under reflected light. While adequate for large, distinct colonies, this approach fails to capture individual organisms or colonies that are transparent, do not reflect light well, or have fine structures, and is unsuitable for high quality timelapses. Darkfield illumination provides superior contrast for organisms on agar surfaces, but existing solutions rely on improvised setups with inconsistent lighting, or on large and expensive benchtop instruments that cannot be housed inside incubators. We present DishCam, an open-source device consisting entirely of 3D-printed parts and inexpensive off-the-shelf electronics, controlled by a Raspberry Pi running custom open-source software. DishCam produces high-contrast darkfield images using angled LED illumination beneath a Petri dish. All components used are off-the-shelf or easily 3D-printed. The camera and lights can be fully controlled through Raspberry Pi using our software. We demonstrate the device across multiple biological systems: Klebsiella aerogenes colonies of differing densities, Trypanosoma brucei colonies doing social motility, Dictyostelium discoideum streaming aggregation, and Caenorhabditis elegans on feeding substrates. The total cost of the basic device is approximately {pound}166 or $222, with the upgraded configuration costing approximately {pound}413 or $552, which includes a high-quality telephoto lens, SSD storage and incubator-compatible cables.
Cocioba, S. S.; Huang, P.-C.; Mallon, J.; Chan, Z.; Geremew, A. W.; Bisson, A.; Kyriakakis, P.
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Here we introduce OpenEvo, a fully open-source, low-cost turbidostat platform for automated continuous culture and directed evolution experiments. Existing tools are expensive, complex, or lack open-source hardware; OpenEvo addresses this gap. OpenEvo is a complete, fully automated evolution platform with detailed, illustrated construction instructions for beginners, open-source software and firmware, and a single device priced around $300. An optional PC-based version offers enhanced functionality, including remote access, programmable evolution cycles, programmable LED stimulation, and a data visualization tool. OpenEvo can cycle through three types of media for positive, negative, and neutral selection conditions, supporting a wide range of experimental designs. We validate the use of OpenEvo by evolving H. volcanii to grow from 15% to 12% salt over ~150 cycles, ~1,000 hours. Evolved cells grew 36% faster than wild-type at 12% salt. Whole-genome sequencing of adapted cells found SNPs and large deletions. We also demonstrate positive and negative selection using the OpenEvo LEDs to drive optogenetics via a Phytochrome B-based optogenetic tool, with light as the selection stimulus during over 4000 hours of growth. OpenEvo lowers the technical and cost barriers for continuous evolution experiments, serves as a teaching tool, and is designed to grow an open community of users who share modifications.
Footer, M. J.; Belliveau, N. M.
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Three-dimensional (3D) printing is increasingly used in cell biology to prototype custom cell culture devices, microscopy chambers, and other experimental hardware, but practical guidance on sterilization and material compatibility remains limited. Here, we develop and validate a low-temperature paraformaldehyde vapor sterilization method for fused filament fabrication (FFF) components that avoids heat-induced deformation. Using bacterial challenge assays with Escherichia coli and Geobacillus stearothermophilus, we confirm effective sterilization of 3D printed components. We also assess the effects of common filaments and adhesives on HL-60 human cell growth to identify materials suitable for cell culture workflows. Most untreated plastics were well tolerated over 48 hours, whereas some formaldehyde-treated materials required post-treatment with ammonia vapor to restore compatibility. Together, these results provide a practical framework for sterilizing and deploying 3D-printed materials in cell culture and biological research. Multidisciplinary Abstract3D printers allow laboratories to quickly make custom experimental equipment, but many printed materials cannot be sterilized using standard methods. We developed a low-temperature sterilization procedure using paraformaldehyde vapor and tested whether common printing materials and adhesives are compatible with cultured human cells. We identified several plastics, silicone-based adhesives and acrylic tapes that can be safely used after sterilization, while others require additional ammonia treatment. These results provide guidance for researchers using 3D printing to build custom laboratory and cell culture devices. Methods SummaryA low-temperature paraformaldehyde vapor sterilization method for 3D-printed FFF components was developed and validated using bacterial challenge assays. Compatibility of common printing filaments, adhesives, and tapes with cell culture workflows was assessed by exposing HL-60 cells to untreated, sterilized, and ammonia- neutralized materials and quantifying cell growth by flow cytometry.
Borch, M. M.; Kehr, P.; Torres, R. A.; Gorter de Vries, P. J.; Larsen, N. J.; Padfield, N.; Nielsen, A. T.
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Gas production and consumption is a direct consequence of microbial activity in environmental and industrial settings. In closed batch cultivations, headspace pressure changes therefore give valuable insights into the microbial metabolism. For laboratory scale anaerobic batch cultivations, manual manometer measurements are routinely applied, as a simple and robust method, but it is labour intensive, causes disturbances in the headspace gas and temperature, leading to suboptimal growth, inhibition and noisy data. We built and tested an automated online pressure sensor for closed batch cultivations. It is designed for microbial cultivation and integrates with sterile and anaerobic cultivation workflows. The system uses an absolute pressure sensor (0-30 bar) mounted on a custom designed PCB, with a gas-tight needle mount. An ESP32 microcontroller logs pressure and temperature locally and generates a Wi-Fi access point for real-time visualization and direct CSV download through a local homepage. We detail hardware and software design decisions, assembly, and validation including long-term stability. Case studies demonstrate the applicability for: a multiphasic biogas kinetics during anaerobic digestion, capturing gas uptake dynamics and metabolic shifts during syngas fermentations and co-feeding experiments, and long-term robustness in a multi-year monitoring of a compressed-air system. More than 130 individual sensors have been deployed over 3 years in laboratories, at various academic and industrial settings. The platform provides reproducible, high-resolution pressure measurements that enable calculation of gas formation/consumption rates and improve experimental throughput without disturbing cultures. Design files, firmware, and example analysis scripts are openly available to support adoption and further development.
Krajnik, B.; Maciejewska, M.; Janeczko, S.; Szczurek, A.
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The queen bee is the central individual responsible for colony establishment, growth, and survival. Reliable confirmation of successful mating, continued queen presence, and normal reproductive performance is essential for effective colony management. We present a queen bee detection system based on an array of Hall-effect sensors and a miniature magnetic tag attached to the queen. The system is designed for continuous operation and real-time monitoring. A prototype was developed, constructed, and evaluated under both laboratory and field conditions. Field experiments conducted in an apiary demonstrated that the system can reliably detect queen bee passages through the hive entrance, enabling the identification of activities associated with mating flights. The results confirm the feasibility of Hall-effect sensing for automated, non-invasive queen bee monitoring and establish magnetic sensing as a promising new measurement modality for precision apiculture.
De Lillo, F.; Smucler, J.
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Electrical stimulation (ES) and transepithelial/transendothelial electrical resistance (TEER) measurements are essential techniques in cell biology and tissue engineering, yet commercial devices for these applications cost between USD 2,500-9,000 and typically offer only one functionality. We present LATEER (Low-cost Arduino-based TEER and Electrical stimulation device), an open-source hardware platform that combines both ES and TEER measurement capabilities at a total cost below USD 100. The device features four independent channels, configurable pulsatile signals (amplitude up to 8.2 V, frequency 0.1-500 Hz, pulse width [≥]0.1 ms), and a resistance measurement range of 300 {Omega} to 1 M{Omega}, with <5% error for R {gtrsim} 4.7 k{Omega}. LATEER uses commercially available graphite pencil leads as electrodes ([~]USD 2 vs. USD 350 for commercial Ag/AgCl electrodes), which demonstrated excellent biocompatibility in cell culture. The system includes 3D-printed electrode holders compatible with standard 12-well and 24-well plates, allowing microscope visualization without electrode removal, and a Python-based graphical user interface for parameter configuration and real-time data acquisition. Because the electrodes remain fixed in the plate lid and only a single cable enters the incubator, both stimulation and resistance measurement can run continuously under standard culture conditions (37 {degrees}C, 5% CO2) without removing the plate or repositioning the electrodes, avoiding the temperature excursions and placement variability inherent to manual chopstick measurements. Validation with human pluripotent stem cell-derived cardiomyocytes demonstrated reliable frequency capture (electrical pacing) of the contracting monolayer, with a capture threshold between 250 and 400 mV/mm and controlled pacing across the 0.5-5 Hz range. TEER functionality was verified with mesenchymal stem cells, where the device resolved cell-density-dependent differences in electrical resistance in real time. All design files, firmware, and software are freely available under the CERN-OHL-S v2 license, enabling replication and customization by research laboratories worldwide. HighlightsO_LIAn open-source device combines electrical stimulation and TEER measurement under $100 C_LIO_LIGraphite electrodes offer biocompatibility at 0.6% cost of commercial alternatives C_LIO_LIFour independent channels with configurable parameters and real-time data logging. C_LIO_LIContinuous run setup in-incubator; no electrode repositioning needed C_LIO_LIValidated with stem cell-derived cardiomyocytes, achieving frequency capture (threshold 250-400 mV/mm) C_LIO_LI3D-printed holders enable microscope visualization without electrode removal C_LI Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=78 SRC="FIGDIR/small/743263v1_ufig1.gif" ALT="Figure 1"> View larger version (28K): org.highwire.dtl.DTLVardef@98f9deorg.highwire.dtl.DTLVardef@13c73aborg.highwire.dtl.DTLVardef@1cdf099org.highwire.dtl.DTLVardef@16ed0cf_HPS_FORMAT_FIGEXP M_FIG C_FIG Specifications Table O_TBL View this table: org.highwire.dtl.DTLVardef@4ef802org.highwire.dtl.DTLVardef@7c651borg.highwire.dtl.DTLVardef@d20013org.highwire.dtl.DTLVardef@102fc89org.highwire.dtl.DTLVardef@111b927_HPS_FORMAT_FIGEXP M_TBL C_TBL
Jaiswal, B.; Black, T.; Namboothiri, H. R.; Pochana, K.; Hu, C. Y.
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Optogenetic control enables light-actuated regulation of gene expression and provides a programmable interface between living cells and electronic systems. However, routine prototyping of optogenetic constructs remains limited by infrastructure. Existing closed-loop platforms often require chemostats, microfluidics, robotic handling, or custom optical sensors, which can increase cost, reduce accessibility, or constrain measurement performance. Here, we present LEMOS 2.0, an updated LED-Embedded Microplate for Optogenetic Studies, a low-cost device for optogenetic stimulation and gene-circuit characterization inside standard off-the-shelf microplate readers. LEMOS 2.0 builds on the original LEMOS platform by increasing throughput from 16 to 32 microwells and reducing light leakage between adjacent microwells, allowing dark conditions to be used as an additional illumination state. The device consists of a 3D-printed frame, individually addressable LEDs positioned next to each microwell, a rechargeable battery, and an onboard microcontroller for Bluetooth-based wireless communication. Biocompatible polydimethylsiloxane microwells are cast directly into the device by replica molding, allowing bacterial cultures to be stimulated while optical density and fluorescence are measured by the microplate reader. This protocol describes the full LEMOS 2.0 workflow, including device fabrication, circuit assembly, Arduino programming, PDMS microwell casting, plate-reader setup, strain and culture preparation, automated experiment execution, device cleanup, and fluorescence/OD600 data analysis. As a demonstration, the protocol uses the CcaSR optogenetic system, in which sfGFP expression is activated by green light and repressed by red light. LEMOS 2.0 is intended to make optogenetic perturbation and gene-expression characterization more accessible to wet-lab users, enabling faster design-build-test-learn cycles without requiring specialized bioreactor or microfluidic infrastructure.
F. Abalde, S.; Bigand, F.; Orciari, L.; Lorini, C.; E. Keller, P.; Parmiggiano, A.; Crepaldi, M.; Novembre, G.
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Joint music making offers an ecologically powerful framework for investigating human social interaction and synchronization. Yet, experimental paradigms often rely on traditional instruments that limit accessibility, reproducibility, and experimental control. In parallel, the use of music for therapy and rehabilitation is expanding, motivating the development of digital musical instruments that can serve research, educational, and clinical purposes. Here, we introduce the e-Music Box Roma (eMB Roma), an open, reproducible digital musical instrument designed to study music making behavior regardless of musical training. The eMB Roma plays preregistered music with tempo controlled by hand rotary movements. Building on the original e-Music Box (Novembre et al., 2015), the eMB Roma retains its intuitive rotary hand control while introducing major innovations: a fully open and 3D-printable design, modular hardware with integrated slider and button controls, polyphonic output with multiple simultaneous instruments, and MIDI compatibility. Additionally, a dedicated graphical user interface allows real-time monitoring, experiment control, device synchronization (like neuroimaging or motion capture devices), and both solo and joint music-making paradigms. The eMB Roma provides a flexible and accessible platform for research contexts, allowing experimental control, reproducibility, and future extensions. Its open design and modularity make it suitable not only for research but also for therapeutic, rehabilitation, and educational applications, where it can support personalized interventions and quantitative assessment of motor performance.
Lodesani, A.; Ross, B. L.; Sridharan, V.; Aiello, C. D.
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Magnetic field effects (MFEs) in biological systems are typically small and experimentally challenging to measure reproducibly across large sample populations. Existing approaches to measure such effects often rely on low-throughput microscopy or custom-built magnetic stimulation systems that provide limited control over magnetic field geometry, synchronization, or experimental automation. Here, we present an open-source magnetofluorescence imaging platform designed for bacterial plate-scale screening of MFEs in live colonies. The instrument integrates a programmable three-axis vector electromagnet, synchronized fluorescence excitation and imaging, and integrated acquisition software with per-frame metadata logging on a hardware-synchronized data acquisition card. An extensive calibration procedure enables accurate generation of arbitrary magnetic field vectors, while synchronized triggering ensures deterministic alignment between field application, illumination, and image acquisition. The system images an entire 100 mm Petri dish in a single acquisition. Typical experiments monitor hundreds of bacterial colonies simultaneously over multi-hour acquisition sequences. Control software, calibration routines, mechanical design files, and acquisition workflows are provided openly to facilitate replication. Instrument performance is demonstrated through detection of magnetic field-dependent fluorescence changes in E. coli expressing the engineered magnetosensitive fluorescent protein MagLOV2. This instrument provides a flexible and scalable platform for high-throughput magnetobiology, synthetic biology, and quantum biology experiments.
Wallerus, A.; Castro e Almeida, S.; Passecker, J.
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A major challenge in behavioral neuroscience is the lack of a unified software framework capable of implementing diverse paradigms across species and experimental setups. Researchers currently face a trade-off: they must either spend significant time developing custom, siloed solutions that hinder reproducibility, or incur substantial costs purchasing inflexible, closed systems. Here, we present Neurokraken, an open-source, Python-native platform designed to overcome these limitations. Neurokraken allows writing experiment progression entirely in standard python, while its core architecture automatically sets up a microcontroller for the connected hardware components and enables python side access with millisecond-precision timing and automatic logging. The system prioritizes ease of use and flexibility, enabling advanced series of events and conditions, the usage of python ecosystem code and packages within experiments, and the addition of any arduino-compatible electronic devices for custom experiments. As a result, users can easily create interactive virtual and real environments to engage, monitor, and record subjects. We present Neurokraken's versatility across a wide range of paradigms, for human and non-human primate psychophysics, and complex rodent behavior in both head-fixed and freely moving paradigms. Its modular design allows for rapid hardware reconfiguration, while a fully customizable user interface enables real-time monitoring and interactive experimental control without compromising timing precision. By uniting laboratory-grade precision with an accessible and flexible open-source philosophy, Neurokraken provides a single, powerful solution to design and execute next-generation behavioral experiments. We hope Neurokraken helps accelerate research, improve reproducibility throughout the neuroscience community, and make advanced behavioral experimentation more accessible through its substantial cost-efficiency.
Cammaert, M.; Wouters, R. I.; van Ede, J. M.; de Hulster, E. A. F.; Mooiman, C. M.; van Dam, P. T. N.; Pabst, M.; van Gulik, W. M.; Daran-Lapujade, P.
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Metabolomics enables the profiling of small-molecule metabolites and thereby captures the biochemical state of a living organism at a given moment and enables to monitor its cellular responses to stimuli. This technique has become a powerful tool in pharmaceutical research, the food industry, and microbial research. Metabolomics aims to obtain an unbiased metabolic profile; however, this is complicated by compound instability, complex and often extensive sample processing, and nonlinear responses in mass spectrometry. Therefore, correcting for metabolite loss and mass spectrometry-related artifacts is essential, typically achieved through relative quantification against an isotopically labelled internal standard for each metabolite of interest. This article describes how to produce 13C-labelled yeast extract and its use as internal standard for metabolomics. More specifically, it provides step-by-step protocols for the fed-batch fermentation, quenching, metabolite extraction, and LC-MS and GC-MS characterization of the internal standard. It also includes a protocol explaining how to use the internal standard for the quantification of metabolites in yeast samples.
Lemke, J.; Spilling, K.
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Sinking marine particles is a key process regulating carbon export through the biological carbon pump, yet direct measurements of sinking dynamics remain limited in many coastal environments. One barrier is that most existing approaches require expensive instrumentation and large research platforms. Here, we present a low-cost, modular method for concentrating fast-sinking particles and measuring their individual sinking velocities under controlled conditions. This combines large settling tanks (110 L) for field-based particle fractionation with a video-based tracking system that quantifies the sinking behavior of natural marine particles. The particle sinking speed chamber is surrounded on three sides by a temperature-controlled water chamber, minimizing the problem of advection during measurements. The post-processing Python script delivers sinking velocity, particle size, circularity, and RGB-based properties for large numbers of particles. The method accuracy was validated using reference beads with known theoretical sinking velocities derived from Stokes law. Field deployments in the Baltic Sea demonstrated successful enrichment of fast-sinking particles and stable operation from both a research vessel and a small boat. Compared to existing methods, the approach substantially reduces logistical and financial barriers while maintaining particle-resolved measurements and compatibility with complementary biogeochemical analyses. This enables a broader observational coverage of sinking particle processes across environments that are currently underrepresented in carbon export studies.
Janarthanan, G.; Chand, R.; Vijayavenkataraman, S.
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Conventional extrusion-based 3D bioprinting encounters limitations in fabricating intricate tissue architectures due to fixed nozzle diameters and fixed deposition orientations. These constraints restrict conformal printing on curved or non-planar surfaces and often necessitate support-intensive fabrication strategies. This work introduces a mechanically simplified extrusion platform inspired by the swivel jet nozzle, featuring a free-degree-of-orientation extrusion head termed the universal extrusion head (Univ-Ex head), coupled with a modular nozzle architecture. The Univ-Ex head employs a swivel-like mechanical design that enables orientation freedom without external actuation in its current implementation, thereby minimizing mechanical complexity while supporting deposition on physiologically relevant, non-planar geometries. Multiple nozzle concepts were developed through comparative CAD iterations, with two representative geometries--a flat nozzle and a conical nozzle--selected for experimental validation. The platform is evaluated through parametric CAD design, stereolithography-printed prototypes, proof-of-concept extrusion experiments, and fluid dynamics simulations performed using FLOW-3D software. Numerical and experimental results demonstrate stable filament formation and clear diameter-dependent extrusion behavior, while simulations further confirm the feasibility of angled and non-planar deposition. A variable-diameter nozzle concept is proposed as a forward design direction to enable real-time adjustment of bioink flow rate and deposition resolution in principle; however, the present study intentionally validates the system using fixed-diameter nozzle variants to maintain stable numerical and experimental boundary conditions. A gear-integrated Univ-Ex head is also presented as a forward upgrade and demonstrated as a single-piece prototype. Collectively, this work establishes a scalable, hardware-focused pathway toward conformal bio-additive manufacturing. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=96 SRC="FIGDIR/small/734010v1_ufig1.gif" ALT="Figure 1"> View larger version (63K): org.highwire.dtl.DTLVardef@8833caorg.highwire.dtl.DTLVardef@33dforg.highwire.dtl.DTLVardef@14d8d11org.highwire.dtl.DTLVardef@685ef0_HPS_FORMAT_FIGEXP M_FIG C_FIG
LIAN, Y.; Zheng, R.; Yang, C.; Luo, L.; Zhang, N.; Lian, G.; Li, B.
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Cystatin-C is an important renal function biomarker, and conventional quantification requires centralized laboratory analyzers, which limits timely testing in primary care and resource-limited settings. To address this need, we developed and validated a simple, rapid, and quantitative smartphone-based (SP) lateral flow immunoassay (LFIA) for measuring serum Cystatin-C. The SP-LFIA platform consists of a colorimetric LFIA strip and a custom SP reader with uniform LED illumination and macro lens for image capture. Quantitative image analysis of the colorimetric signal is performed by a dedicated application using a pre-defined third order polynomial calibration model. Following systematic optimization, the assay demonstrated a wide quantitative range of 0.32-8.00 mg/L, with a limit of detection of 0.15 mg/L. Analytical validation conducted according to CLSI guidelines showed excellent precision, with intra- and inter-assay coefficients of variation below 10%, and no significant interference from bilirubin, triglycerides, hemoglobin, or rheumatoid factor. Accelerated stability testing confirmed robust strip performance after storage at 50 {degrees}C for 28 days. Method comparison using 100 clinical serum samples showed high agreement with a commercial PETIA reference method (R{superscript 2} = 0.993) and minimal bias. These results indicate that the developed smartphone-based LFIA provides a reliable, cost-effective, and practical tool for point-of-care Cystatin-C monitoring.
Khoroshun, E. V.; Kozlov, V. A.; Ivanov, I. V.; Momynaliev, K.
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BackgroundContinuous glucose monitoring (CGM) systems are used not only for retrospective assessment of the glycemic profile but also for real-time decision-making, including automated insulin delivery. Accordingly, CGM performance characterization must capture not only the agreement of individual paired values but also the systems ability to reproduce the direction, rate, amplitude, and shape of glucose concentration change. Summary metrics, most notably MARD, cannot establish whether an observed deviation reflects an error in the formation of the test profile itself, a constant sensor offset, amplitude compression, a change in response rate, temporal misalignment, or hysteresis. ObjectiveTo adapt a programmable flow-based in vitro platform for the separate assessment of the experimentally delivered glucose profile and the dynamic response of CGM systems, and to propose a set of metrics that decomposes dynamic error into its components. MethodsGLU profiles were generated by programmable mixing of solutions at a constant total flow rate of 2 mL/min. Actual GLU concentration was independently measured with a SUPER GL2 glucose analyzer. Four static levels, three repeats of a 5.5[->]12.0[->]5.5 mmol/L profile, three repeats of a 6.0[->]3.0[->]6.0 mmol/L hypoglycemic profile, three 5.0[->]15.0[->]5.0 mmol/L profiles at different rates, one complex 4[->]18[->]3[->]12[->]5.5 mmol/L profile, and two proof-of-concept sensor experiments at 100- and 200-min transitions were investigated. Dynamic response was characterized by bias, MAE, RMSE, MARD, amplitude transfer coefficient K_A, rate transfer coefficients K_up and K_down, normalized shape RMSE, residual shift, and hysteresis loop area. ResultsAt the static levels, measured GLU exceeded the programmed value by 0.234-0.780 mmol/L. In the repeated 5.5[->]12.0[->]5.5 profiles, the ratio of actual to programmed rate was 0.978-1.083 on the rising phase and 0.987-1.157 on the falling phase, while the amplitude transfer coefficient was 0.967-1.066. In the hypoglycemic profile, minimum GLU was 2.55- 2.96 mmol/L, and time below 3.0 mmol/L was 15.2-72.6 min. The measured rates of 0.0519, 0.1045, and 0.2027 mmol/L/min preserved the intended ratio of approximately 1:2:4. In the complex profile, the programmed plateau of 18 mmol/L was not reached: mean measured GLU was 16.20 mmol/L. For CGM-A, K_A was 0.682 and 0.650, and K_up/K_down were 0.666/0.730 and 0.634/0.626; the corresponding values for CGM-B were 1.228 and 1.128, and 1.564/1.328 and 1.276/1.145. Hysteresis loop area differed 5- to 10-fold between the two sensor responses, exceeding an order of magnitude at the 100-min transition. ConclusionThe programmed concentration should be treated as a control setpoint, rather than as a reference measurement. The "programmed trajectory -- measured glucose -- CGM output" cascade first allows quantitative assessment of the agreement between the programmed and actually realized profile and only then separate characterization of sensor response. Decomposition of dynamic error into amplitude, rate, shape, and hysteresis components reveals differences that a single MARD value or correlation coefficient cannot capture.
De Keyzer, L.; Deserranno, K.; Skevin, S.; Van Hoofstat, D.; Deforce, D.; Van Nieuwerburgh, F.
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Recombinase polymerase amplification (RPA) enables rapid nucleic acid testing in low-resource environments, but poorly characterized byproducts can compromise assay specificity and cause false-positive results. Here, we amplified the thirteen original CODIS core loci and Amelogenin to characterize recurrent RPA artefacts and establish conditions that reduce their formation. First, RPA products were analyzed for two reference samples by Oxford Nanopore Technologies sequencing. This revealed two distinct classes of multimeric products: primer multimers and amplicon multimers, consisting of repeated primer or amplicon sequences, respectively. Individual artefacts contained up to 281 primer copies or 22 amplicon copies, demonstrating the extensive range of these products. Next, we performed an optimization study to evaluate the effects of reaction temperature and reagent concentrations at two representative loci, D3S1358 and D5S818. Among the conditions tested, temperature had the most pronounced effect. Reducing the temperature from 42{degrees}C to 34{degrees}C increased the relative target amplicon fraction from 15% to 83% for D3S1358 and from 84% to 98% for D5S818, while maintaining or increasing absolute target concentration. Lower primer concentrations and higher T4 UvsX concentrations also reduced multimer formation, although lower primer concentrations reduced target yield and caused allelic dropout. Finally, amplification at 34{degrees}C was evaluated across all fourteen loci by sequencing. Relative to 42{degrees}C, the target read fraction increased by more than 5 percentage points for 7/14 loci in one reference sample and 9/14 loci in the other, with the largest improvements at multimer-prone loci. These findings identify multimers as an important class of RPA artefacts and establish reaction temperature and T4 UvsX concentration as promising conditions to improve RPA specificity.
Gaidica, M.; Rosengart, M.
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Light reaching the retina is a primary regulator of human circadian physiology, acting largely through melanopsin-expressing retinal ganglion cells with peak short-wavelength sensitivity. Delivering known, repeatable retinal doses outside the laboratory is difficult because conventional light sources leave viewing geometry, gaze, and ambient conditions uncontrolled. Consumer extended-reality (XR) glasses fix a bright binocular display in constant geometry relative to the eye, but their suitability as calibrated photic stimulators has not been established. Here we validate a commercial micro-OLED XR display (VITURE Luma Ultra) for controlled retinal photostimulation. A purpose-built host application renders exact 8-bit RGB stimuli while independently controlling hardware brightness and logging all intensity-determining state; spectral radiance was measured at the retinal position of a 3D-printed phantom head with an open-source miniature spectroradiometer, anchored to absolute units by a luminance transfer calibration. The blue primary peaks at 461 nm (FWHM 43 nm), is spectrally invariant across a >10-fold intensity range, and at maximum output delivers an estimated 299 lx melanopic equivalent daylight illuminance, above consensus daytime recommendations, while remaining roughly two orders of magnitude below photobiological safety limits. The red primary is visually effective with minimal melanopic drive (melanopic DER 0.10), enabling spectrally shifted evening stimulation. Unlike the immersive virtual-reality headsets previously used for calibrated light delivery, the see-through form factor preserves the wearer's view of the surroundings--relevant for clinical monitoring in supervised settings such as the intensive care unit. These results show that consumer XR glasses can serve as a dose-calibrated platform for wearable photostimulation using an open-source measurement chain, and provide groundwork for application-layer dose-response studies.
Caira, T.; Tokihiro, J.; Shaposhnikov, A.; Whitten, J. M.; Su, X.; Shin, A.; Robertson, I. H.; Nicholson, T. M.; Olanrewaju, A. O.; Berthier, E.; Theberge, A. B.; Berthier, J.
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Control of fluids is a hallmark of microfluidic systems and fundamental for the successful application of microfluidic devices. Trigger valves use geometric features to autonomously control the release of fluids in microfluidic devices. Our previous work has adapted geometries used in closed trigger valve systems to enable use in open systems, allowing for open microfluidic devices with up to three trigger valves. Here, we focus on the parallel co-flows produced by sequential release of trigger valves and present a model that predicts their layer widths as a function of the geometric characteristics of the different side channels of each trigger valve. We show layered co-flows with widths as low as 50 microns. Additionally, we expand the use of trigger valves in open microfluidic devices by incorporating 1) varied step heights, 2) devices with up to seven trigger valves, and 3) use of varied fluids and plastics. To validate the implementation and use of these trigger valves in open systems, we have developed a theoretical framework to compare predicted outcomes (i.e., fluid travel distance, velocity, and layering width) with our experimental values. This theoretical work offers applications in various fields, including hydrogel patterning for 3D cell culture, organ-on-a-chip models, at-home sample preparation, and autonomous microfluidic systems for biosensing.
Sommer, S.; Dhmine, O.; Mateos Langerak, J.; Dobbie, I. M.
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Microscopes are essential tools for discoveries on a scale invisible to the unaided human eye. The development of immuno-fluorescence followed by molecular biology techniques and fluorescent fusion proteins have revolutionised the use of optical microscopy in bioscience. The quality of the data produced is dependent upon the sample, its preparation and the instrument used. However, instruments can degrade over time without easily visible changes to the produced images and, in turn, negatively impacts results. By testing instruments and doing comparisons between results over time and between different instruments, problems can be highlighted and corrective action can be taken. Using small fluorescent beads the point spread function (PSF) of the microscope can be recorded and the image resolution measured. Beads were prepared in a concentration matched to the field of view size and dried onto coverslips and mounted on slides. The beads were then imaged as 3D Z-stacks of sufficient size to fully enclose the PSF of the system. This data was uploaded to OMERO and processed using OMERO-metrics, an OMERO plugin developed for this purpose. This paper summarizes the development of workflows and protocols to enable this process, presents the results obtained and demonstrates the detection of significant instrument issues.
Esmaeilzadeh, K.; Hosseini, M.; Etghani, S. A.; Vahabie, A.; Yekani, M.
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Low-cost and open-source neural recording systems are increasingly important for expanding access to electrophysiological research. However, many existing platforms still rely on specialized hardware or limited modularity, restricting flexibility for laboratories seeking customizable solutions. Here, we developed and evaluated a modular neural recording platform constructed entirely from commercially available components. Recordings were compared against the ground truth. The platform successfully recovered local field potential (LFP)-like waveforms in most conditions and detected spike-like activity during direct connection recordings. Principal component analysis and k-means clustering further demonstrated the ability to distinguish multiple simulated spike waveforms. Signal quality varied across configurations, with saline recordings and preamplifier integration introducing increased noise and reduced detectability. These findings demonstrate the feasibility of building affordable and modular electrophysiology systems using widely accessible hardware. Although the current implementation has limitations in sampling rate, noise performance, and in vivo validation, the presented framework provides a practical foundation for future customizable open-source neural recording.