Biosensors

Traditional sample preparation for flow cytometry is often labor-intensive, operator-dependent, and reagent-consuming, limiting its suitability for automated and point-of-care biosensing applications. To address these challenges, this study presents a functional modular microfluidic system integrating immunomagnetic beads (IMBs) to enable automated intracellular immunofluorescence (IF) staining. The modular microfluidic platform is enabled by a dynamically actuated three-dimensional magnetic field that couples with IMBs within a microfluidic reaction chamber, requiring only one-dimensional magnet translation to induce effective three-dimensional bead motion. This magnetic–chip cooperative strategy significantly enhances microscale mixing and cell capture, facilitating automated immunostaining of the radiation biomarker in CD4⁺ cells. Finite element simulations were employed to guide magnetic field design by analyzing magnetic force distributions and identifying key parameters, including magnet material, size, spatial arrangement, and chip–magnet distance. Experimental validation using CD4⁺ cell capture confirmed the effectiveness of the magnetic mixing strategy, revealing ∇B·B as the critical design parameter. An N52 NdFeB magnet (6 mm diameter, 10 mm height) positioned within 2.2 mm of the chamber centerline stably retained IMBs at flow rates below 200 µL/min. Under optimized conditions (magnet translation speed of 8 mm/s and a 15 min mixing duration), a maximum cell capture efficiency of 86% was achieved. Subsequent automated γH2AX IF staining demonstrated a strong linear dose–response relationship (R² > 0.9) in mean fluorescence intensity. This study demonstrates a robust and scalable strategy for automating complex IF staining workflows, highlighting the potential of magnetic-field-assisted microfluidic platforms for biosensing applications requiring reliable intracellular biomarker detection.

Correct focal positioning is essential for microscopy imaging of live moving subjects such as Caenorhabditis elegans. However, many methods can be too slow to perform real-time control to keep the subject in focus. In this work, we propose a convolutional neural network-based method to perform one-shot prediction of the optimal focusing distance, without the need to scan iteratively the optical axis to find the optimal position. A new data augmentation technique is proposed, and its effectiveness is validated through statistical analysis. This technique is shown to improve results without the need for additional data collection. Several architectures are trained in z-stacks of images, using the proposed data augmentation technique, and compared on a validation set. Through this comparison, we find that the ConvNext V2, a novel architecture in this context, outperforms other models proposed in previous works. Furthermore, the impact of the Field of View used for the model’s prediction is studied, with the aim of further understanding the influence of spatial resolution and spatial compression on the performance of the model.

Avian influenza (AI), particularly highly pathogenic avian influenza (HPAI), represents a serious and growing threat to global poultry production, international trade, and human health security. Control of AI is complicated by the high evolutionary rate of influenza A viruses, which drives antigenic diversity and ongoing emergence of novel strains. Effective surveillance and disease management therefore depend on timely and accurate diagnostics. While conventional methods—including virus isolation, reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and enzyme-linked immunosorbent assays (ELISAs)—remain effective and widely used, they are limited by long turnaround times, the need for specialized equipment, and reliance on highly trained personnel. In addition, strict state and federal regulatory requirements restrict testing to a limited number of authorized laboratories. Although these regulations are essential for maintaining diagnostic accuracy and quality assurance, they place substantial strain on laboratory capacity during outbreaks and delay actionable results. The need for rapid, on-site decision making has driven interest in alternative diagnostic approaches, including biosensor technologies. A major limitation of current diagnostic strategies is the lack of robust DIVA (Differentiating Infected from Vaccinated Animals) capability. In countries such as the United States, where poultry vaccination against AI is not routinely practiced, the absence of DIVA-compatible diagnostics has hindered adoption of vaccination as a disease management tool, as seropositive birds and products face significant trade restrictions. Biosensor platforms capable of enabling DIVA strategies offer a potential pathway to support vaccination while preserving surveillance integrity. This review examines the current landscape of AI and HPAI diagnostics, emphasizing the limitations of traditional approaches and the opportunities presented by biosensor platforms. We evaluate electrochemical, optical, piezoelectric, and nucleic-acid-based biosensors, with particular attention to biorecognition strategies, performance metrics, field deployability, and applications supporting subtype discrimination, DIVA implementation, and One Health surveillance.