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Microfluidics and lab-on-a-chip technologies have redefined fluid manipulation at small scales, providing powerful solutions for challenges in biophysics, biotechnology, and disease diagnostics. This talk will highlight advancements in microfluidic biosensing through a few examples, showcasing their transformative potential in healthcare and biomedical research. An optomicrofluidic device utilizing localized surface plasmon resonance (LSPR) with gold nanospikes enables SARS-CoV-2 antibody detection in plasma within 30 minutes at a detection limit of ~0.5 pM (0.08 ng/mL). A complementary duplex electrochemical immunosensor further distinguishes infection-induced from vaccine-induced antibodies in under 7 minutes, supporting large-scale sero-surveillance efforts. Expanding beyond infectious disease diagnostics, a multiplexed opto-microfluidic platform facilitates high-sensitivity prostate cancer biomarker detection, while a surface-imprinted polymer system enables selective and rapid bacterial detection. Additionally, wearable paper-based microfluidic devices provide real-time pH and sodium ion monitoring in sweat, offering non-invasive tools for personalized health tracking. Our results highlight the versatility of microfluidics in advancing diagnostics, personalized medicine, and environmental monitoring, delivering scalable, rapid, and precise solutions for complex biomedical challenges.

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Wound healing is an essential biological process for maintaining homeostasis and, ultimately, for survival. We investigate the mechanisms underlying extreme wound healing in Stentor coeruleus, a single-celled organism, capable of recovering from drastic membrane wounds exceeding half of the cell surface. This talk focuses on our recent effort on developing a microfluidic platform for the manipulation and reproducible wounding of the cell. We demonstrate a microfluidic "guillotine" for bisecting cells in a continuous flow, and a "SMORES" platform to immobilize the cells for laser ablation. We further discuss the extension of the tool for dissecting live tumors to generate organoids that preserve the tumor immune microenvironment for applications including the testing of immunotherapy on a chip.

Microfluidics provides a "toolbox" for the study of biological systems, probing the very length, time and force scales that are relevant in biology. A mechanically important entity of a biological cell is the cytoskeleton, an intricate network of protein filaments - actin filaments, microtubules and intermediate filaments. These biopolymers, which all have very different properties in terms of bending stiffness, stretchability and polymerization dynamics, are complemented by passive cross-linkers and active molecular motors. Our research aims at designing experiments that lead to a better understanding of the mechanical and dynamic properties of the cytoskeleton and, in particular, allow us to quantify our results. I will give examples of studies on pure or mixed networks using continuous flow and droplet microfluidics.

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Liquid biopsy offers a minimally invasive alternative to tissue biopsy, providing crucial insights for personalized treatment through circulating biomarkers. However, current methods face challenges including high cost, complexity, and limited sensitivity. We present innovative microfluidic solutions to address these limitations. Our "lab-on-a-chip" systems efficiently analyze disease biomarkers in biological fluids, with a particular focus on extracellular vesicles (EVs) for cancer diagnostics. We introduce EV-CLIP, a droplet-based digital method that revolutionizes EV RNA profiling. This technology employs charged liposomes (CLIPs) for the fusion with EVs, achieving exceptional sensitivity in detecting EV-derived miRNAs and mRNAs. Using just 20 µL of plasma without prior EV isolation, EV-CLIP demonstrated remarkable accuracy in detecting EGFR mutations (L858R: AUC 1.0000; T790M: AUC 0.9784) across 83 patient samples. Its capability for serial monitoring during chemotherapy enables precise tracking of rare EV subpopulations, advancing our understanding of EV heterogeneity in disease progression. Additionally, we present a hand-powered centrifugal device for bacterial isolation and antibiotic susceptibility testing, designed specifically for resource-limited settings. These microfluidic innovations collectively aim to accelerate the clinical implementation of liquid biopsy technologies, ultimately enhancing patient care through improved accessibility and diagnostic accuracy.

Nanoparticle Tracking Analysis (NTA) has emerged as a fast and vital characterization technology for Extracellular Vesicles (EVs), Exosomes and other biological material in the size range from 30 nm to 1 μm. While classic NTA scatter operation feeds back the size and total particle concentration, the user typically cannot discriminate whether the particle is a vesicle, protein aggregate, cellular trash or an inorganic precipitate. The fluorescence detection capabilities of f-NTA however enables the user to gain specific biochemical information for phenotyping of all kinds of vesicles and viruses. Alignment-free switching between excitation wavelengths and measurement modes (scatter and fluorescence) allow quantification of biomarker ratios such as the tetraspanins (CD63, CD81 and CD9) within minutes. Furthermore, specific colocalization studies using c-NTA gives a deeper understanding of the composition of biomarker on single particle.

Colorectal cancer (CRC) as the second leading cause of global cancer deaths poses critical challenges in clinical settings. Cancer-derived small extracellular vesicles (sEVs), which are secreted by cancer cells, have been shown to mediate tumor development, invasion, and even metastasis, and have thus received increasing attention for the development of cancer diagnostic or therapeutic platforms. In this talk, I will introduce the sEV-targeted systematic evolution of ligands by exponential enrichment (E-SELEX) to generate a DNA aptamer (CCE-10F) that recognizes and binds to CRC-derived sEVs. This novel aptamer possesses high binding affinity and specificity for CRC-derived sEVs, effectively discriminating CRC-derived sEVs from those derived from normal colon cell, human serum, and other cancer cells. Furthermore, the aptamer significantly suppresses the critical processes of metastasis, including cellular migration, invasion, and angiogenesis, which are originally induced by sEVs themselves. These findings are highly encouraging for the potential use of the aptamer in sEV-based diagnostic and therapeutic applications.

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Over the last 18 years, Kloé company developed a complete range of equipment dedicated to UV lithography applications, in perfect agreement with the microfabrication requirements in Microfluidics. Thus, Kloé company did the bet, in the early 2000, that the development of researches and industry in Microfluidics would rapidly grow. So that, over the same time, Kloé company continuously followed and exchanged with the Microfluidics community to first well understand and then anticipate its needs in terms of microfabrication techniques and performance, in order to enable fabricating from simple to more demanding microfluidic chips like Lab on a Chip / Organ on a Chip.

Among a very large range of 12 different machines, covering from soft lithography / masking systems to very high resolution direct laser writers particularly suitable for fast prototyping, high aspect ratio as well as thick layers laser processing, Kloe introduces one of its latest innovations that is Dilase3D : a 3D-Printer specifically developped to meet the expectations for 3D-printing in Microfluidics.
Typically elaborated from the specifications of researches in Microfluidics and Medical Sciences, that were looking for one tool enabling to both fabricate large volume pieces, but still with very high-resolution patterning capabilities, this equipment also demonstrated more recently its capability to combine different materials for the fabrication of one piece/object, that multiplies its capabilities to fabricate very demanding and ever more complex microchips/microstructures. This way, we ensure our partners to benefit from the one of the most performing and cost effective 3D-printing solution in that domain, in agreement with their expected level of performance and their available budget.

Precision medicine has changed the healthcare paradigm by introducing molecular diagnosis techniques that, combined with liquid biopsy, make non-invasive personalised diagnosis possible. However, the wide implementation of these techniques in clinical routine is far from reality.

Microfluidics has demonstrated numerous advantages for isolation and characterization of liquid biopsy biomarkers in oncology. These systems can be coupled with embedded biosensors to quantify disease biomarkers with increased sensitivity and throughput, enabling their implementation in clinical routine.

In this talk, I present our most recent work in the development of liquid biopsy assays for integrated isolation and analysis of multiple circulating biomarkers and their validation in clinical settings.

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Bacterial Extracellular Vesicles (BEVs) are small vesicles naturally released by Gram-negative and Gram-positive bacteria during their growth process into the surrounding environment. These vesicles are composed of BEVs components, lipopolysaccharides, proteins, nucleic acids, and more, playing a significant role in the physiology and pathogenesis of bacteria. Here is an introduction to the role of BEVs in disease diagnosis and treatment:
Pathogenicity and Transmission: BEVs vesicles can carry bacterial virulence factors and genetic material, which can be transmitted to host cells, thereby promoting infection and disease occurrence.
Immunomodulation: BEVs contain a variety of bacterial antigens that can stimulate the host's immune system, making them candidates for vaccine development to induce an immune response against bacteria.
Disease Diagnosis: Since BEVs contain specific bacterial components, they can serve as biomarkers for early disease diagnosis and monitoring.
Therapeutic Applications: BEVs vesicles can be used as drug carriers to deliver medication directly to the site of infection, improving therapeutic efficacy and reducing side effects. Studies have shown that engineered BEVs can be used in cancer immunotherapy by activating or enhancing the host's immune response to tumor cells.
Research Challenges and Future Directions: Future research may focus on how to precisely control the composition of BEVs and how to use them to develop more effective diagnostic tools and treatment methods.
The study of BEVs vesicles is an interdisciplinary field involving microbiology, immunology, molecular biology, and nanotechnology, among others, and their application prospects in disease diagnosis and treatment are broad.

Introducing EXODUS, our innovative automatic exosome isolation system, which employs nanofiltration techniques combining the double-coupled harmonic oscillation and periodic negative pressure oscillation. This unique integration ensures the high yield and purity of label-free exosome isolation. Due to its capbility of processing diverse range of samples, EXODUS has been widely embraced in research for disease biomarker discovery. Besides, EXODUS also features a large-scale model, EXODUS-T, specifically tailored to meet the demands of industrial-scale production.

Extracellular vesicles (EVs) have been recognized as the ideal alternative for drug delivery and therapeutic development. EVs are secreted from live cells with minimal toxicity and immunogenicity to in vivo systems, and enable the delivery of a variety of large size payloads including CRISPR Cas9/sgRNA RNP complexes. Recent discoveries also showed the strong ability of EVs to cross difficult biological barriers. However, the heterogeneity of EV surface presentation and diverse molecular makeup make them difficulty to study. We reported novel molecular engineering and microfluidic approaches for surface modification, intravesicular payload loading, and biomarker discovery. Particularly, the CRISPR gene editing, and AI directed extracellular vesicle surface engineering overcome the current challenge in tissue targeting specificity in the field, which could lead the promise for developing next-generation precision therapeutics and gene therapy.

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Our study identifies Amyloid Precursor-Like Protein 1 (APLP1) as a novel biomarker within brain-derived extracellular vesicles (EVs) in peripheral blood, offering significant potential for the early detection of neurodegenerative diseases. Rigorous analysis confirmed APLP1+ EVs as cerebrally originated through unique small RNA expression patterns and validation using transgenic Thy-1 GFP M line mice, where GFP and APLP1 co-localized in hippocampal neuron-derived EVs. These findings highlight APLP1+ EVs as promising non-invasive diagnostic markers, providing a transformative approach to understanding and detecting neurodegenerative disease progression.

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In today's fast-paced scientific landscape, transitioning from microfluidic prototyping to commercial-scale injection molding presents unique challenges. This talk will delve into the intricacies of moving from commonly used prototyping techniques—3D printing, MEMS fabrication, PDMS casting, and CNC machining—to full-scale production. Each method offers specific hurdles that need careful consideration to ensure a seamless transition to injection molded polymer cartridges. We will explore tailored strategies to address these challenges, offering solutions to streamline the process and highlighting alternative approaches when direct transitions prove difficult. Our focus will be on practical solutions that enhance scalability and maintain the integrity of the original prototype's design and functionality. Join us to uncover the keys to efficient and effective transformation from prototype to product in the microfluidic domain, setting a new standard for innovation in manufacturing.

Multi-stimulus responsive soft materials with integrated functionalities are elementary blocks for building soft intelligent systems, but their rational design remains challenging. In this report, we introduce an intelligent soft architecture sensitized by magnetized liquid metal droplets that are dispersed in a highly stretchable elastomer network. The supercooled liquid metal droplets serve as microscopic latent heat reservoirs, and their controllable solidification releases localized thermal energy/information flows for enabling programmable visualization and display. This allows the perception of a variety of information-encoded contact (mechanical pressing, stretching, and torsion) and noncontact (magnetic field) stimuli as well as the visualization of dynamic phase transition and stress evolution processes, via thermal and/or thermochromic imaging. The liquid metal-elastomer architecture offers a generic platform for designing soft intelligent sensing, display, and information encryption systems.

Hepatocellular carcinoma (HCC) is a major malignancy worldwide and is particularly prevalent in Mainland China and Hong Kong. Metastasis is the main attribute of high mortality rate and recurrence rate in HCC patients. Tangible evidence reveals a conducive tumor microenvironment for metastasizing cells is of paramount importance during the process of metastasis. Extracellular vesicles (EV) that comprises a heterogeneous population of membranous vesicles has become the crucial mediator of intercellular communication. The expanded diversity of EVs has great potential to embrace more mechanisms underlying functions of EVs in HCC. In this talk, the functional role and clinical relevance of small extracellular vesicle (exosome) and exomere in HCC will be discussed. Our study unraveled the mechanisms through which EV contribute to the progression of HCC, particularly in promoting tumor growth, angiogenesis, and metastasis, which are crucial processes in cancer development. Furthermore, we explored mechanisms of EV internalization into recipient cells and EV biogenesis. The insights gained from these studies suggest the potential of targeting EVs as a therapeutic strategy for HCC.

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