Projects

Reproduction is regulated by the endocrine system and its disturbances by endocrine disruptive chemicals (EDCs) may lead to infertility. As humans are constantly exposed to EDCs through the use of common household items and personal care products, it is important to test chemicals for their potential activity as endocrine disruptors affecting reproductive function. Project MERLON aims to study the effects of EDCs on sexual development and function in order to deliver new approach methodologies (NAMs) for EDC identification. While MERLON targets the vulnerable stages of development from fetal to puberty, MERLON2, with additional partner TalTech, will add one more sensitive window of susceptibility in female reproduction to the project: the adult preovulatory ovarian follicle, where the oocyte maturation takes place. In collaboration with TalTech, it was recently demonstrated that follicular somatic cells (FSCs) lose sensitivity to follicle stimulating hormone (FSH) in the presence of a mixture of 13 EDCs. FSH is crucial for both, the oocyte maturation and for the synthesis of steroid hormones by the FSCs. We have also demonstrated the intricate heterogeneity of somatic cells in the ovarian follicle. The roles that FSC subpopulations play in the adverse effects of EDCs is unknown and unaddressed by the initial MERLON project. MERLON2 will complement the aims of the consortium by developing NAMs based on single cell transcriptomics, automated image analysis and machine learning to understand the effect of EDCs on FSC subpopulations and their sensitivity to FSH. This will increase the research output for MERLON in the number of proposed NAMs and quantitative adverse outcome pathways. As a result of MERLON2 the range of stakeholders will enlarge, increasing the public awareness related to the harmful health effects of EDCs, and proposing new approaches to resolve the complicates issue of testing substances in everyday products for their adverse effects on human fertility.

The overall objective of the original CHIRALFORCE project is to demonstrate enantiomer separation in a compact, on-chip, photonic platform that is fabricated using standard silicon-based technology. This CHIRALFORCE2 hop-on project enhances the original project by providing automated in-line platform for the analysis of chiral separation for this CHIRALFORCE photonic chip. Separation of enantiomers from mixtures is essential, especially in early phase drug discovery processes when many mixtures need to be separated. CHIRALFORCE aims to revolutionize the field of chiral chemistry by introducing a radically new strategy for separating enantiomers by using chiral optical forces in silicon-based photonic integrated waveguides to separate enantiomers. The successful implementation of CHIRALFORCE project (development of separator chip) relies on fast and accurate feedback on the enantiomer separation. However, current state-of-the art technologies for checking the enantiomer separation: e.g. circular dichroism (CD) spectroscopy or High-Performance Liquid Chromatography (HPLC) lack off-the shelf capabilities for rapid in-line separation monitoring that is needed in CHIRALFORCE project. CHIRALFORCE2 addresses this need by providing a platform for in-line monitoring of the chiral separation down-stream from the CHIRALFORCE separator chip. We use interdisciplinary approach combining automation, electronics, optics and IT disciplines. The monitoring of in-line chiral separation will be achieved by CD-spectrometry or absorbance detection depending on the microfluidic and optical requirements from CHIRALFORCE project. Both scenarios are supported by designated software for the signal analysis and feedback.

New or reoccurring bacterial threats are a major challenge of this century, and a delayed response due to the lack of field-testing options risks human lives and causing an epidemic. Classical microbiology techniques are relatively slow, while cytometric methods allow the measurement of cell count, morphology etc. in an easy, reliable, and fast way. State of the art flow cytometers are high-throughput benchtop instruments that are neither portable nor cheap enough for field testing, causing logistic delays in bacterial testing in remote areas and conflict zones or where infrastructure is limited. The goal of this R&D activity is to create the proof of concept of and develop the methodology for low-cost, fully portable flow cytometers based on droplet microfluidics, which will not only allow field analysis of bacteria, but will have a single-cell resolution. Furthermore, through cognitive electronics, the system will be easy to use and fully automated from sample input to result output.

The most significant results of this project include developing a user-friendly tool for microbiology studies. This tool was used to figure out how growth of individual bacteria cells varies during exposure to different metals. This vital knowledge will contribute to ongoing experiments about the possibility of metals affecting individual bacteria cells’ ability to grow in the presence of antibiotics, and to overall research in antibacterial substances and antibiotic resistance. The newly developed tool has also opened the door for future experiments and new collaboration projects both regionally and internationally. Researchers and students alike, no matter their educational or financial background, will be able to apply the tool for their own research topic. Through this project the PI has obtained valuable insight into leadership, experimental procedures, networking, and writing which has contributed to development into an independent researcher.

There are many antimicrobial substances (e.g. antibiotics) in the use globally that are starting to lose their activity against pathogenic microbes (antimicrobial resistance). This is serious threat to human health and economy in general. This project aimed to develop new experimental technologies to investigate certain molecular mechanisms in the nature that can lead to such resistance. We used novel droplet technologies to investigate such mechanisms at single cell level in bacteria population. Water-in-oil-droplets are like small test-tubes that enable carrying out parallel investigation of biological and chemical phenomena in tens and even hundreds of thousands of such “test-tubes”. This high-throughput approach helps understanding biological problems better as large experimental datasets help seeing the patterns better with more confidence. In our case we investigated phenotypic heterogeneity in genetically identical bacteria populations that can lead to survival of bacteria during antibiotic treatment. Firstly, we developed user-friendly droplet analysis tools that help investigating biological experiments in droplets. Using droplets for experimental analysis is not yet mainstream, often because of the need for highly specialized tools or trained personnel. We addressed this issue by developing and comparing droplet tools that are easy to implement in non-droplet biological and chemical laboratories worldwide. Secondly, we used developed tools in our own laboratory to investigate how different is antibiotic impact on bacteria that are in different stages in their life cycle. This knowledge helps understanding why some bacteria manage to survive during antibiotic treatment without developing mutations. This in turn can help researchers to prevent spread of antibiotic resistance. As a project manager I am extremely satisfied that we developed user-friendly droplet technologies that help widening the access to droplet technologies worldwide.