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Our research centers on the study of microfluidic lab-on-a-chip devices applied to environmental analysis, biomedical research and the synthesis of smart materials. Microfluidic devices offer the possibility for in-the-field and point-of-care analysis provided the devices are portable, require only minimal external instrumentation and little power and are robust. In our group, we are investigating a developing paper microfluidic devices for on-site measurement of water quality markers that are being tested by members of the general public who upload data via a custom-built app for data collection with high spatial and temporal resolution. Furthermore, we develop on-site workflows that enable farmers in resource-limited settings to monitor soil chemistry on their own land.

The discovery and validation of circulating biomarkers for minimally invasive diagnostics of complex diseases have been a major focus of AIT’s Molecular Diagnostics group. Multi-omics approaches for biomarker development from both tissue and cell-free body fluids, including the utilization of EV-derived miRNAs, have been successfully established. In this presentation, the extensive multi-omics technology workflows developed by our research group will be introduced and selected results from a study conducted on patients with coronary artery disease will be highlighted, demonstrating the power of integrating various omics layers to identify and validate circulating biomarkers. Additionally, diverse EV biomarker studies will be reported, showcasing research across a range of diseases, including metabolic, immune-mediated, and cardiovascular diseases. EVs derived from different biofluids, such as serum, plasma, cell-free saliva, and cerebrospinal fluid, have been utilized, demonstrating the versatility and promise of EVs in disease diagnostics.

Presentation of the challenges faced by astronauts during long-duration space missions and how the European Space Agency (ESA) is supporting their health and performance through research on the impact of space conditions on the human body and the development of countermeasures and point of care technologies to investigate changes and mitigate those consequences. The presentation will focus on ESA's SciSpacE team, which conducts research in ground analogues, suborbital platforms, and on the International Space Station (ISS), and how the research conducted in space can benefit terrestrial medicine. The process of ideation, call for proposals, review, selection, and implementation of research projects will also be outlined. The presentation aims to provide insights into how ESA is working to keep astronauts healthy and productive during long-term space missions and how these efforts can also benefit medical science on Earth.

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Lab-on-a-Chip devices can enhance the study of disease-causing microorganisms and tackle their effects on global health. We describe the design, manufacture and use of devices use to study the forces these organisms use to to infect their hosts.

Here, we present the development of a BPM sensor to measure cytokines, exemplified with the detection of Interleukin-6 and TNF-alpha using various coupling strategies. Furthermore, approaches for continuous sampling will be shown.

Immunology has advanced significantly in recent years thanks to a better knowledge of molecular protein recognition, which has allowed for the creation of innovative treatment options such as genetic vaccines and cell therapies. However, there is a need to accelerate the process of epitope recognition by proteins in order to generate personalized therapies. ElectroMed has been a project with six European partners in which we created a concept for a programmable peptide screening platform based on an array of Immuno-FET sensors with programmable peptide receptors synthesized in-situ using electrochemically generated acid.

A proof-of-concept has been demonstrated by controlling the deprotection of acid labile groups, which enables a high purity synthesis of crude peptides. We remove diffusion-limited processes in chemistry by automating the peptide synthesis and using microfluidic flow in-situ synthesis in planar sensors, allowing us to expedite time to results. FET sensors have the potential to provide online quality control of peptide synthesis by detecting the fingerprints of acid labile group protection and deprotection, as well as superior information quality because label-free sensors can provide kinetic information.

In conclusion, our approach has the potential to speed research into peptide protein screening of linear peptide sequences. In this talk we will review our results and provide the context for the applications in immunology.

Effective intracellular delivery of biomolecules is crucial in analyzing and engineering cell functions, for applications in therapeutic development, diagnostics, and drug delivery towards personalized medicine. Traditional bulk cell culture models employed so far for intracellular delivery, overlook cell population heterogeneity, while single-cell patterning offers robust statistical analyses without compromising cellular variability. In this work, a high-throughput single-cell patterning approach using microcontact printing is employed, where a micro-pillar stamp is used to imprint distinct proteins and finally pattern single-cell to small clusters of cells depending on the micro-pillar diameters. This approach is universal for any protein-cell combination and has 97-99 % patterning efficiency. This single-cell patterning was used as a platform for massively parallel optoporation-mediated intracellular delivery of small to very large biomolecules such as PI dye (668 Da), Dextran (3000 MW), SiRNA (20-24 bp), and enzyme (464 kDa). An easy-to-fabricate and simple-to-use 2D array of titanium micro-dish device is used to facilitate near infrared mediated optoporation. which disrupts the cell plasma membrane, allowing biomolecules to enter into cells with high efficiency (~96±2%) and cell viability (~98±1%). Our compact, robust, and simple-to-print approach has potential for single-cell therapy and diagnostics research, providing high specificity and sub-cellular accuracy in single-cell level biological processes.

Extracellular vesicles (EVs) are generated by various cell types, including endothelial cells. The presence of EVs in fetal bovine serum (FBS), commonly used in cell culture, has been recognized as a potential confounding factor. To elucidate this, human brain microvascular endothelial cells (HBMEC) were cultured for 2 or 24 hours in the presence of EV-depleted FBS (EVdS). Cell viability, gene and protein expression, and EVs isolated from these cells were assessed. Additionally, EV uptake, ICAM-1 expression, and monocyte adhesion to HBMEC exposed to EVs were also examined. Elevated apoptosis rates in cells cultured with EVdS for 2 and 24 hours was observed. Within 2 hours, there was an upregulation of IL8, followed by the downregulation of IL6 and IL8 after 24 hours. Proteomic analysis revealed that EVs cultured for 2 hours (EV2h) were enriched in proteins associated with ribosomes and carbon metabolism, while those cultured for 24 hours (EV24h) exhibited proteins linked to cell adhesion and platelet activation. Moreover, HBMECs exposed to EV2h displayed increased ICAM-1 expression and monocyte adhesion compared to cells exposed to EV24h. These findings highlight that HBMECs cultured with EVdS produce EVs with distinct physical characteristics and protein content that varies across time.

This paper presents the optical bunching of protein microcrystals in a 3D printed microfluidic device for improving the sample delivery for serial crystallography studies at X-ray free electron lasers.

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