This training course is being taught by the ALine team, including Dr Stefano Begoloo, a veteran in the microfluidics, system design and product development space and by Antonio (Tony) Pineda. The goal of this training course is to present an actionable set of tools for translating microfluidic design into products.

Specifically, the following topics will be addressed:

1. Looking at product development from the view point of your stakeholder

2. Create milestones with deliverables that de-risk and build momentum

3. Benchmark your results to assure data integrity; good science is necessary for a viable product

4. When to transition from a working alpha to a beta prototype

5. Understand and balance function and complexity to avoid a high cost of manufacture

The overall goal of this training course is to engage with fellow attendees, engage with the instructor and discuss topics as they relate to your product development efforts.

Subsequent to this course is the 2.5-day main conference and a deep dive into these areas during the conference in presentations and discussions.

**If you are involved in microfluidic product development, this training course is a must-attend**

This short course offers a practical introduction to the manufacturing of microfluidic devices, focusing on the transition from lab-scale prototyping to high-volume production.

Topics will include:

1. Key materials used in microfluidics: polymers, silicon, and glass

2. Prototyping techniques such as 3D printing, CNC micromachining, laser cutting, soft lithography (PDMS), and MEMS-based fabrication—along with their capabilities, limitations, and typical applications

3. Challenges in scaling up, including design adaptation, material compatibility, and process scalability

4. Production-ready solutions, including micro-injection molding, various chip bonding strategies, PDMS scale-up methods, and considerations for manufacturing microfluidic MEMS devices as distinct from conventional MEMS used in sensors and actuators

The course will be illustrated with real-world industry case studies, showcasing the latest trends shaping the future of microfluidics in diagnostics, life sciences, and pharmaceutical applications.

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As a person’s physiological regulation of biomarker molecules is challenged by acute illness, exposure to toxins or even surgery, the concentration these molecules can give important information about their health. For premature infants their regulation systems have yet to mature, so they can also suffer rapid changes in biomarker concentrations that can have devastating health consequences for the baby. Our view is that to monitor such biomarker changes effectively ideally requires moment-by-moment measurement of blood or tissue concentrations. The person acts as their own control allowing acute deterioration to be noticed quickly.

We have been developing a range of sensing and biosensing solutions for the invasive, minimally invasive, and non-invasive monitoring of people in healthcare situations. Microfluidics coupled to novel biosensors provide a valuable means of clinical sampling and robust quantification of measured biomarkers.

I will describe the key challenges in the development of such integrated microfluidic sensing devices and present our recent data from the neonatal intensive care unit amongst other projects.

Detection of molecular biomarkers in point-of-care settings requires an automated workflow that begins with a clinical sample, processes it with reagents for a selective assay, and delivers an accurate quantitative output in less than 30 minutes, while utilizing low-cost disposable sensors and a portable inexpensive instrument.

This presentation will summarize three recent multidisciplinary and collaborative efforts towards this goal.

First, we will describe the “PathTracker” system (in collaboration with Rashid Bashir, Enrique Valera and Bill King at UIUC), in which a plastic cartridge (produced by either injection molding or additive manufacturing) processes a droplet of whole blood to release the nucleic acid content of viral pathogens that are selectively detected by multiplexed loop-mediated isothermal amplification (LAMP) assays for Zika, Dengue, and Chikungunya virus. The PathTracker cartridge provides precisely metered volumes off all reagents from onboard reservoirs, while pneumatic pressure for mixing is provided by milliliterrotating a knob.

Second, we will describe a “Biosensor Integrated Recovery Device” (BIRD) that pushes milliliter-scale plasma volumes through a series of nanoporous filters to extract and concentrate nano-objects (such as viruses or extracellular vesicles) within a specific size range, followed by exposure of the filtrate to an integrated photonic crystal biosensor.

Finally, we will share recent efforts to develop and demonstrate paper-based lateral flow test strips that utilize ultra bright plasmonic fluor tags that can be detected with a pocket-sized, fluorescence reader. The system has been characterized for high sensitivity detection of gonorrhea antigen in urine.

What are the areas that must be considered to carry a microfluidic product brainchild from brilliant concept to benchtop model, miniaturised prototype, beta and eventually launched product? There are many, and several should be considered in parallel to give you a fighting chance of making it to product launch on the other side of the gap ... before your funds run out or your competition steals your lunch!

Based on 13+ years of consulting to start-ups and multinationals alike (and occasionally to university tech transfer offices and public tech support agencies) on a variety of product development, IP review and due diligence projects, a survey of these PD scope areas and a general outline of how these projects typically proceed will be provided. Topic areas that will be addressed include product definition, IP landscaping, quality and regulatory standards, product concept and materials considerations, iterative design, prototyping, testing & evaluation, and transfer to manufacturing.

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We have developed a multi-parametric high-throughput and high-sensitivity flow-based method for counting single molecules, and applied this method to the analysis of individual extracelluar vesicles and particles (EVPs). EVPs are promising biomarkers but they are highly heterogeneous and comprise a diverse set of surface proteins as well as intra-vesicular cargoes. Yet, current approaches to the study of EVPs lack the necessary sensitivity and precision to fully characterize and understand the make-up and the distribution of various EVP subpopulations that may be present. Digital flow cytometry (dFC) provides single-fluorophore sensitivity and enables multiparameter characterization of EVPs, including sin­gle-EVP phenotyping, the absolute quantitation of EVP concentrations, and biomarker copy numbers. dFC has a broad range of applications, from analysis of single EVPs such as exosomes or RNA-binding proteins to characterization of therapeutic lipid nano­particles, viruses, and proteins. dFC also provides absolute quantita­tion of non-EVP samples such as dyes, beads, and Ab-dye conjugates.

Managing seemingly incompatible performance metrics such as sensitivity, size, speed, complexity, and versatility has long been a challenge for biomedical sensors, often resulting in prioritization of some aspects over others depending on the application. I will discuss the integration of optical and electrical single molecule analysis in a lab-on-chip format and their use in a wide variety of practical applications. By integrating optical waveguides and solid-state nanopores in a microfluidic setting, target-agnostic detection with high sensitivity, low complexity, rapid turnaround and wide dynamic range was demonstrated. Examples will include infectious disease diagnostics, liquid biopsies for cancer treatment, continuous exosome analysis in 3D organoid tissue culture platforms, and on-chip spectroscopy as a new avenue for lab-on-chip miniaturization.

Despite a large number of publications, electrowetting-on-dielectric (EWOD) digital microfluidics (DMF) is yet to achieve the once-anticipated wide adoption. To democratize DMF for easy adoption, a group of labs has been developing a cloud-based platform of standardized design and manufacturing. The platform is aimed to empower any user to design, obtain, and operate DMF chips (https://edroplets.org). For chip design, a web-based EWOD chip design platform allows layout rules and automated wire routing. For chip manufacturing, a web-based EWOD chip manufacturing platform facilitates fabrication of four types of EWOD chips (i.e., glass, paper, PCB, and TFT) to suggest the foundry service workflow of the future. For chip control, a compact EWOD control system is available along with web-based operating software. Our goal is to inspire academic and commercial stakeholders to join the initiative toward a DMF ecosystem for the masses.

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3D printing enables the direct fabrication of complex, enclosed microfluidic devices, eliminating the need for molds, layer alignment, and post-fabrication bonding that characterize many conventional processes. However, considerable confusion persists regarding the feasibility of 3D printing for the interconnected negative features essential to microfluidics. This often stems from equating manufacturer-reported resolution specifications with actual achievable dimensions, as well as from insufficient understanding of the underlying processes and optimal strategies for their application. This presentation introduces a conceptual framework to clarify what is possible and how to effectively leverage 3D printing for microfluidic device fabrication, including both DLP and LCD resin printers. We also emphasize the expanded capabilities afforded by full user control over the printing process. As illustrative examples, we present results from our custom DLP 3D printers and resins, including channels with 2 μm × 2 μm cross-sections, valves featuring 15 μm × 15 μm active areas, integrated dose-response assay chips, and a single device incorporating 11,200 valves with 100% yield that demonstrates the complexity now becoming feasible with tailored 3D printing processes.

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Trapping, linearization, and imaging of single molecule DNA is of broad interest to biophysicists and has applications to nucleic acid analysis methods such as karyotyping and optical mapping. In this work, we drive DNA molecules with an axial electric field through microchannels and study the dynamics on a single-molecule basis. We discovered a new method to trap and visualize DNA molecules. Namely, above a threshold electric field, individual DNA molecules become pinned to the channel walls at a vertex and fold into a hairpin shape as they are stretched in the direction opposite to the electric field. We hypothesize that polyvinylpyrrolidone (PVP), a neutral polymer added to the buffer to suppress electroosmosis, adsorbs on the channel surfaces and forms a scaffold onto which DNA becomes entangled. Upon removal of the electric field, DNA molecules undergo relaxation within a few seconds to a Brownian coil around the vertex. After 10’s of seconds, DNA is released and free to diffuse and electromigrate. We analyze the conditions needed for trapping, the relaxation dynamics, and the repeatability of vertex pinning. The method enables high quality imaging of single-molecule DNA with high throughput using simple-to-fabricate fluidic structures.

The rise of personalized medicine is reshaping traditional healthcare, enabling predictive analytics and tailored treatment strategies. In this talk, I will discuss our progress in developing wearable, implantable, and ingestible electrochemical biosensors for real-time molecular analysis. These bioelectronic systems autonomously access and sample diverse body fluids—including sweat, interstitial fluid, gastrointestinal fluid, wound exudate, and exhaled breath condensate—enabling continuous monitoring of key biomarkers such as metabolites, nutrients, hormones, proteins, and drugs during various activities. To facilitate scalable, cost-effective manufacturing of these high-performance, nanomaterial-based sensors, we employ laser engraving, inkjet printing, and 3D printing techniques. The clinical utility of our biosensors is being evaluated in human and animal studies, focusing on applications such as stress and mental health assessment, precision nutrition, chronic disease management, and personalized drug monitoring. Additionally, I will highlight our efforts in energy harvesting from both the body and the environment, opening the door to battery-free, wireless biosensing technologies. By integrating electrochemical biosensing with advanced bioelectronics, we aim to revolutionize personalized healthcare, offering new possibilities for diagnostics, continuous monitoring, and therapeutic interventions.

Digital microfluidics offers a more integrated and cost-effective approach to next-generation sequencing (NGS) sample preparation by enabling discrete control of liquid droplets at the nL–µL scale. To bring this technology into practical use in real-world laboratories, key technical challenges had to be addressed. This presentation also introduces a next-level platform that builds on the strengths of digital microfluidics while significantly extending its capabilities. Designed for full automation, enhanced robustness, and scalability, the platform integrates essential NGS sample preparation steps into a compact, cost-effective consumable. The result is a practical, high-performance solution that simplifies workflows and supports high-throughput molecular biology applications.

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In the rapidly evolving field of microfluidics, the journey from concept to production often presents a fragmented and time-consuming challenge. This talk presents an integrated workflow designed to streamline microfluidic innovation, from digital design to physical device, in record time. We will introduce a fully web-based CAD and simulation tool tailored for microfluidics, enabling users to iterate faster and more accurately. Next, we explore a true 3D mold fabrication solution that bypasses the limitations of traditional lithography. Finally, we present Flexdym, a next-generation polymer that merges the processability of thermoplastics with the flexibility of silicone, enabling rapid prototyping and scalable manufacturing. By combining digital tools, novel materials, and modular fabrication systems, we offer a new paradigm for microfluidic design, validation, and production.

Scaling up microfluidic innovations from laboratory prototypes to large-scale manufacturing presents unique technical and operational challenges. This presentation addresses critical considerations for transitioning microfluidic designs from proof-of-concept devices to mass-produced commercial products. Leveraging Hicomp's expertise in microfluidic chip prototyping, fabrication, and process optimization, we will discuss practical strategies for selecting suitable materials, optimizing chip designs for manufacturability, ensuring robust quality control, and streamlining production workflows. Real-world examples and insights will illustrate common pitfalls, effective solutions, and best practices to facilitate successful scale-up. This session aims to provide attendees with actionable guidance on bridging the gap between laboratory innovation and high-volume commercial fabrication.

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Micro- and nanoplastics are emerging environmental and biomedical concerns, with growing interest in their detection and health impacts. This talk presents our recent work on lab-on-a-chip platforms for the sampling and characterization of plastics across environmental and biological matrices. I will introduce microfluidic particle-trapping and microfiltration devices integrated with Raman spectroscopy and machine learning for rapid identification of micro- and nanoplastics in water samples, along with an acoustic approach for separating microplastics from diluted blood samples. Finally, I will discuss in vitro studies on the cellular responses to polystyrene particles, emphasizing the effects of environmental weathering and simulated gastric fluid exposure on particle surface properties and cytotoxicity.

The advancement of automated lab-on-a-chip systems often relies on custom-formulated 3D printing resins, a practice that creates a significant barrier to widespread adoption. To address this, we investigated the "off-label" use of widely available commercial dental resins as a cost-effective alternative for fabricating monolithic microfluidic devices with integrated active components. We explored several valve architectures and, after iterative design optimization, identified geometries that could be reliably fabricated. We then characterized their performance and demonstrated valve utility in a practical application of an on-chip peristaltic pump. The results show comparable performance to custom resins at lower expense and error risk, enabling accessible microfluidics.

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While great advances in medicine has been made in the past century, the overall infrastructure of the healthcare system has not progressed. Patients are still expected to travel to a centralized location for discrete, reactionary based care where the healthcare provider only has a brief window to assess the patient’s health. Unless the symptoms are overt at the time of examination, the subjective evaluation relies heavily on the self-reporting of symptoms from the patient. This often results in delayed or improper diagnoses. In contrast, we know that physiological signals precede clinical deterioration. We have developed a suite of soft, low-cost, unobtrusive, Band-Aid © like physiological sensors to continuously monitor patients’ cardiovascular and pulmonary functions. We seek to continuously quantify subtle physiological changes to predict – and eventually prevent -- the onset of acute clinical events.

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Join this interactive workshop to experience a complete microfluidics prototyping workflow, from digital design to a real-world device, in under an hour. Participants will use FLUI'DEVICE, a browser-based platform built for intuitive microfluidic design and simulation. Each design will be brought to life using Flexdym, an alternative to PDMS, and the Sublym hot embossing system, allowing rapid chip prototyping with thermal bonding, no cleanroom required. The session also includes through-hole embossing, showcasing new possibilities in chip functionality and multilayer integration. Whether you're new to microfluidics or looking to accelerate your workflow, this hands-on session is designed to inspire and equip you with practical tools and materials to bridge the gap between design and device.

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In vitro diagnostic tests represent an important application area for microfluidics and even nanofluidics. However, these tests can be complex because of the number of processing steps that must be employed for successfully carrying out each assay, the diverse range of diseases that must be tested each requiring a different workflow, and the diverse set of tests that must be undertaken (screening → diagnostic → prognostic → response to therapy → recurrence). To complicate the process of bringing new tests to the clinical area is the rigorous validation that must be undertaken both analytically and clinically, and the FDA approval process required for all in vitro diagnostic tests.

To address the aforementioned issues microfluidics and even nanofluidics provide an excellent platform for in vitro diagnostic tests and there are examples already in the commercial sector that provide excellent examples, such as Next Generation Sequencing and their flow cell for nucleic acid analyses, and point-of-care tests for infectious disease testing a good example being the RT-qPCR systems for COVD-19.

In this workshop, I will discuss various aspects of the different testing formats for in vitro diagnostics all of which will have a microfluidic thematic focus. Various topical items that will be discussed include:

• Market size and needs for in vitro diagnostic testing
• The do’s and don’ts of bringing new testing platforms to the market (the fall of Theranos)
• Analytical and clinical figures-of-merit for in vitro diagnostic tests
• Requirements for FDA approval
• The use of liquid biopsy markers and microfluidics for in vitro diagnostic testing
• Requirements of microfluidic and even nanofluidic systems for diagnostic testing
• Integrated microfluidic systems for carrying out multi-step assays for diagnostics
• Practical examples:

o Infectious disease testing (COVID-19)
o Point-of-care diagnostic tests for cerebral vascular diseases
o Microfluidics and nanofluidics for the management of cancer-related diseases

As technologies mature, they are incorporated into product applications, and as applications mature, standards often evolve around them. The development of standards serves the purposes of consumer convenience and user/bystander safety, and they can be driven by industry and supported by governments. Examples in science include oscilloscopes and power supplies that use BNC and banana plug connectors, manual or robotic lab pipettors for sample preparation that adopt the 9-mm spacing standards for 96-well microtiter plates, and fluidic connections that use Luer lock connectors.

The purpose of this workshop is to look at the evolution of standardised approaches in microfluidics products, product development, and research. The increased use of standards in microfluidics is an important step, as it may facilitate widespread acceptance by removing barriers such as incompatible connectors, inconsistent technical terminology, unsuitable materials, and unquantified or uncharacterized figures of merit.

Our workshop speakers will tell us about what their organisations do, and how standardisation in their operations facilitates the development of microfluidic products by drawing from a library of standard microfluidic components or processes. This, in turn, leads to faster and more predictable development stage, which ultimately benefits their customers, and the microfluidic community at large.

**This interactive workshop is open to all registered conference attendees**

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Neurodegenerative diseases are a growing global health challenge, with current treatments obstructed by difficulties in modeling, diagnosing, and treating complex conditions. This research addresses one of the key obstacles—understanding the blood-brain barrier (BBB)—by developing a novel microfluidic system that mimics the dynamic environment of the BBB. This work focuses on the design of an acoustofluidic device that integrates a surface acoustic wave (SAW) driven micropump to generate pulsatile flow and shear stress within a vascular-like lumen. The micropump is used in conjunction with a closed-loop system to drive fluid through the microfluidic device, which is created to mimic the BBB model. Central to the design is the investigation of an acoustic whispering gallery mode (WGM) that can manipulate cell behavior in the vascular-like wall, with the potential to temporarily disrupt the BBB and enable drug delivery. Computer-aided design (CAD) software and finite element analysis (FEA) simulations were used to optimize the microchannel system, with promising results for both fluid flow generation and WGM frequency tuning. Both devices were fabricated using the stereolithography technique for mold printing employing a combination of two types of resins to create a complex 3D microchannel system. Microfluidics were examined using a CT X-ray scan and subjected to testing. Theoretical and experimental results from the research would be discussed.

Early, accurate, and accessible detection of infectious RNA is pivotal for effective communicable disease intervention and public health surveillance. However, current RNA detection methods, such as RT-qPCR, require thermal cycling and bulky centralized infrastructure, limiting their utility in decentralized or resource-limited settings. We present a high-throughput droplet digital CRISPR-Cas13a assay that enables ultrasensitive, isothermal, and rapid quantification of HIV-1 RNA. Leveraging a millipede-based optofluidic platform capable of generating and analyzing ~15 million droplets per minute, this approach integrates single-RNA droplet partitioning with CRISPR-Cas13a-mediated collateral cleavage for fluorescent signal amplification. Digital quantification across droplets enables qPCR-level sensitivity without the need for thermal cycling. We focus on refining the assay’s sensitivity and specificity by optimizing CRISPR RNA design, Cas13a ortholog selection, and reaction conditions to mitigate false-positive activity and maximize signal-to-noise in picoliter-scale droplets. These efforts aim to establish a robust proof of concept for a droplet digital CRISPR-Cas13a platform that can support both clinical RNA screening and real-time infectious disease monitoring. Beyond clinical diagnostics, we envisage broader applications in areas such as outbreak surveillance, wastewater monitoring, and food safety, where point-of-care-compatible and ultrasensitive RNA quantification can enhance frontline response and biosafety infrastructure.