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Blood is a complex liquid, and the way it circulates in the human body is linked to its rheological properties. The high deformability of red blood cells plays a major role in hemodynamics, as do the shape and complexity of the vascular networks. We are developing microfluidic vascular networks that attempt to mimic as closely as possible the great complexity of microcirculation, so as to be able to study under the microscope the way in which blood circulates in them.

Resistive Pulse Sensing (RPS) is a label-free and single-molecule detection approach that requires simple instrumentation to implement and as such, can be mobilized to be integrated into in vitro diagnostic assays for not only detecting but identifying key disease-associated biomarkers with high analytical sensitivity. Thus, RPS is a logical choice for coupling with liquid biopsy markers for the precision management of a variety of diseases due to the significant mass limits imposed on any assay in which a liquid biopsy marker is used. We have developed a unique measurement modality and sensor technology (dual in-plane nanopore sensor) that couples RPS to nanoscale electrophoresis, which has recently garnered attention due to unique molecular-dependent information it can provide. Not does it generate the typical RPS measurement parameters, but also the molecular-dependent electrophoretic mobility, which we call the time-of-flight (ToF). The RPS parameters coupled with the ToF and machine learning leads to high detection efficiency and classification accuracy of single molecules harvested from liquid biopsy markers, including nucleic acids and proteins. Our devices, which are made from plastics via high-scale production modalities (injection molding), consist of channels with dimensions ranging from 1 to 100 nm (effective diameter) that are 10’s of microns in length. In this talk, I will discuss the operational parameters and unique applications of our dual in-plane nanopore sensor for three compelling applications: (1) determining the fill status (empty versus full) of adeno-associated viruses (AAVs), which serve as carriers of gene therapy drugs; (2) peptide fingerprinting of single protein molecules; and (3) DNA/RNA single-molecule sequencing.

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In recent years, advanced in vitro pulmonary platforms have witnessed exciting developments pushing beyond traditional preclinical cell cultures at the air-liquid interface. Here, I will discuss ongoing developments that aim to deliver new generations of anatomically- and physiologically-inspired lung-on-chip platforms that coincide with the sprouting of human-relevant preclinical in vitro models. Motivated by translational endpoints. I will exemplify efforts covering inhalation-based inflammation and infection of the airways to novel strategies for improved aerosol delivery to the lungs. Lastly, we will explore the integration of induced pluripotent stem cell (iPSC) derived airway epithelial cells into lung-on-chips.

The basis of my talk is polymer microarray technology which has been developed in the Bradley group.
In my talk I will introduce polymer microarray technology and describe how this approach has been used in a number of applications – ranging from sensor optimisation/immobilisation to the provision of cellular substrates/scaffolds for a variety of screening applications. Specifically I will cover:

The use of polymer microarray technology to discover substrates with optimal binding and responsiveness of fluorescent reporters and their immobilisation onto the ends of optical fibres for measuring pH around tumours

The use of polymer microarray technology to discover polymers that bind cancer stem cells and control their differentiation) (with Tetsuya Tega, Stem Cells, 2016) with application in cancer screening.

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Harnessing 3D printing for microfluidic device fabrication is challenged by the need for sub-100 μm features, especially the negative space structures central to microfluidics. This presentation showcases custom Digital Light Processing (DLP) 3D printers and optimized materials developed to tackle these demands and push the boundaries of resolution. We introduce a photopolymerization simulation tool to explore the enhanced parameter space created by our generalized 3D printing method. This method expands the available parameter space to achieve negative feature resolutions approaching the printer’s physical limits. We demonstrate the application of these innovations to high-resolution passive structures (e.g., channels, mixers) and active components (e.g., valves, pumps), integrated into functional devices such as dose-response assays and cell chemotaxis chips. Example achievements include channels with 2 μm x 2 μm cross-sections and valves with 15 μm x 15 μm active areas, highlighting the potential of these innovations to redefine microfluidic fabrication.

Magnetic Resonance Imaging (MRI) uses hydrogen nuclei in strong magnetic fields to non-invasively sense tissue or structural properties. The interaction between the hydrogen nuclei and distortions of the magnetic field form an essential source of imaging information in clinical MRI. 3D micro-printing enables to manufacture features that create well-controlled magnetic field inhomogeneities in the micrometer scale. This enables a novel test bed to study and develop MRI methods with microstructural sensitivity.

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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The detection of ultra-rare variants in liquid biopsy samples is often hindered by the highly fragmented nature and low concentrations of cell-free (cf) DNA and circulating tumor (ct) DNA. In non-advanced, non-metastatic cancers, ctDNA constitutes only 1–2% of total cfDNA, making its extraction and analysis particularly challenging. However, current DNA extraction and purification methods are limited by high costs, complex instrumentation requiring routine maintenance, and lengthy processes that reduce DNA recovery and compromise integrity for downstream applications such as PCR and sequencing. To address this critical gap, our team has developed a patented DNA extraction and isolation (DEI) membrane and assay, designed to efficiently capture and retain more DNA from liquid biopsy samples. Our technology enhances DNA yield and integrity, offering a cost-effective and scalable solution for integration into point-of-care diagnostic systems, including microfluidic platforms, laboratory diagnostic systems, and PCR-based workflows. Here we have demonstrated a microfluidic-based digital PCR platform where the DEI membrane was integrated into our current platform towards effective and efficient quantification of viral load.

This presentation tries to cover technical, manufacturing, and legal aspects in the process of bringing a very early stage device to the market and avoiding costly mistakes upfront.

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This study develops a long-term circulating tumor cell (CTC)-derived model from a triple-negative breast cancer (TNBC) patient who has developed resistance to standard treatments. CTCs are isolated from blood using a negative enrichment approach and cultured in a basement membrane extract with organoid medium. The resulting organoids proliferate for more than 10 passages, maintain TNBC characteristics, and show high resistance to platinum-based therapy, mirroring the patient’s tumor. Drug screening identifies potential therapeutic vulnerabilities, highlighting the model’s relevance for studying metastasis, therapy resistance, and immune interactions in TNBC. This preclinical tool facilitates the development of targeted therapies and a better understanding of CTC biology.

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PP TechSales represents the innovative dispensing technology of PolyPico Technologies Ltd., which simplifies the dispensing process for R&D, small-scale, and fully automated production. Using patented disposable dispensing cartridges (DCs) eliminates cross-contamination, minimizes maintenance, and reduces costs associated with traditional dispensing methods. With the unique precision, PolyPico’s dispensers can generate pico-, nano-, or low microliter volumes at speeds of up to 10,000 droplets per second. Thanks to dispensers’ versatility and multiple DC orifice sizes there are supported various solutions, including DMSO, buffers, oils, glues, and even 100% glycerol. This cutting-edge technology delivers unique accuracy and efficiency, eliminates intensive washing, buffer consumption or carry-over risks, while enabling recovery of unused samples.

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We explored the development of a novel integrated optofluidic system for application like UV-Vis spectroscopy analyte detection. Total Internal Reflection (TIR) approach requires a cladding material with a refractive index (RI) lower than the fluidic core (eg. water, n=1.33). The silica aerogel was chosen as a highly nano-porous material whose RI can be tuned from 1.05 to 2.2 using a thermal densification process. Simple channels were shaped using PET polymer wires inserted during the gelification step and dissolved under the supercritical drying one. After hydrophobic treatment, we built an aerogel-based optofluidic (OFAB) modules, including a manifold system able to integrate both optics and fluidics. First, we studied the detection of multiple concentrations of the dye ’Brilliant Blue FCF’ at 630nm as a function of sample length and channel diameter. Then OFAB approach was used to detect a range of drug concentrations (Ropivacaine), diluted in a biologically relevant medium (PBS) at 263nm.

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Nanoscale systems for drug delivery have received considerable attention over the past decades, as they have the potential to overcome limits associated with conventional drug therapies, providing a solution to medical challenges that urgently require new therapeutic approaches. Nanodrugs can improve the therapeutic index of the loaded drug by providing protection against degradation, enabling controlled release and distribution, and increasing bioavailability. Advanced functionalization strategies have been used to confer them long-circulating properties and facilitate targeting to specific cells. These efforts have led to the introduction into clinical practice of a few nanoscale systems for tumor therapy. However, their small number, compared to the variety of promising systems proposed, reveals a considerable gap between favorable preclinical results and real clinical performance. Biological barriers, inherent to tumors and their microenvironments, pose formidable challenges. To bridge this gap, our focus turns to three-dimensional (3D) culture methodologies. Unlike conventional 2D cultures, 3D models better replicate the heterogeneity, pathophysiology, and structural architecture of real tumors. Our ongoing efforts involve constructing heterotypic multicellular 3D tumor spheroids, serving as robust screening tools. This approach aims to enhance our understanding of nanodrugs penetration and accumulation within the tumor mass. By identifying key parameters, we strive to maximize therapeutic benefits.

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The presentation will focus on the following topics:

Introduction on High-Grade Gliomas (HGGs) and its Tumor microenvironment

To properly study and challenge HGG Tumor microenvironment (TME) is important to recapitulate several aspects: (e.g., brain extracellular matrix, cell-to-cell interactions)

Bioreactor-based culture of neurospheroids: from neural cell differentiation to incorporation of microglia-derived cells: from high batches to miniaturization

HGG 3D cell culture establishment
Inclusion of HGG cells within neurospheroids: building up of immunosuppressive TME and invasive phenotype

Conclusion and future perspectives

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Intestinal barrier dysfunction has been associated with various non-gastrointestinal diseases common in aging populations, emphasizing the need to investigate age-related changes in gut function. While impaired barrier integrity is a hallmark of gastrointestinal disorders, the direct impact of aging in humans remains unclear. To address this, we developed an in vitro gut-on-a-chip platform incorporating an impedance sensor to assess senescence-driven barrier alterations. This platform, equipped with membrane-based electrode arrays, enables real-time, non-invasive monitoring of epithelial barrier function. In order to generate a senescent gut model, Caco-2 cells were treated with 0.8 μg/mL doxorubicin (DXR), a known senescence inducer, for six days. Our findings reveal DXR-induced impedance increases, cell hypertrophy, and upregulation of senescence markers p21 and CCL2, confirming a senescent phenotype. By covering approximately 57% of the cultivation area, the integrated electrodes allow reliable and continuous assessment of barrier dynamics. This sensor-integrated gut-on-a-chip system provides a valuable tool for studying age-related intestinal dysfunction and advancing our understanding of barrier integrity in aging populations.

The rheological properties of biological tissues are crucial in wound healing or morphogenesis. We have developed an innovative microfluidic device which enables studying in depth the rheology of model tissues by realizing micropipette aspiration of ~20 spheroids (3D cell aggregates) in parallel. Thanks to dynamic stimulation, we show that the coupling of fluid flow inside the aggregate with its deformation (poroelasticity) needs to been taken in account to properly describe the tissue response: permeability contributes to the transmission of stress, followed by cell deformation and finally cell reorganization in the tissue. Cell scale imaging and immunostaining are also used to assess the relationship between microstructure and rheological properties. Finally, the approach can be extended to other tissues. In collaboration with a team specialized in embryogenesis, we are assessing spatial heterogeneities in cail embryos, in order to relate cell fate and differentiation to biophysical properties.

Precision medicine promises to transform diagnostics and acute care improving patient outcomes and reducing healthcare costs. Through accurate patient stratification a real opportunity to impact patient care can be realized. Pena et al. (EBioMedicine 2014. 1(1):64-71) previously published the discovery/validation of an 31-gene transcriptional signature predicting sepsis development. The gene expression signature of immune cell reprogramming was subsequently narrowed using machine learning resulting in a new 6-gene classifier compared to 2 housekeeping genes (SepsetER). Herein, we developed a molecular assay for SepsetER, that predicts risk of clinical deterioration within the first 24 hours in patients with prospective sepsis. A stand-alone centrifugal microfluidic instrument that integrates the entire workflow for detection of the SepsetER classifier in 50µL of whole blood was developed and tested. The analytical process was split between two microfluidic cartridges that when connected enable RNA isolation and ddPCR biomarker detection to be performed in sequence automatically. This PREcision meDIcine for CriTical care (PREDICT) system was validated using an independent cohort of label-free patients with suspected sepsis (N=30). The PREDICT system had a high sensitivity of 92%, specificity of 89%, and an overall accuracy of 88% in identifying the risk of imminent clinical deterioration in patients with suspected sepsis.

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