Ms. Alexandra Tsouka Profile

Alexandra Tsouka

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Proceedings Article | 4 March 2019 Paper

Automated laser-assisted synthesis of microarrays for infectious disease research

G. Paris, J. Heidepriem, A. Tsouka, M. Mende, S. Eickelmann, F. Loeffler

Proceedings Volume 10875, 108750C (2019) https://doi.org/10.1117/12.2516781

KEYWORDS: Glasses, Polymers, Microscopes, Magnesium, Laser systems engineering, Luminescence, Cameras, Laser processing, Image processing, Calibration

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We developed a next-generation method for chemical in–situ combinatorial biomolecule array synthesis. This allows for an unprecedented combinatorial freedom in the automated chemical synthesis of molecule arrays with very high spot densities. Key feature of this new method is an automated positioning and laser transfer process: Small solid material spots are rapidly transferred from a donor film to an acceptor surface, requiring only minute amounts of materials. The transfer is performed with different and easy-to-produce donor slides. Each donor slide bears a thin polymer film, embedding one type of monomer. The coupling reaction occurs in a separate heating step, where the matrix becomes viscous and building blocks can diffuse within the material and couple to the acceptor surface. Since these transferred material spots are only several nanometers thin, this method allows for a consecutive multi-layer material deposition of e.g. activation reagents and amino acids. Subsequent heat-induced mixing facilitates an in–situ activation and coupling of the monomers. Positioning several of such resin spots, containing different chemical reagents, on top of each other, will enable for the first time in such small dimensions unique chemical synthesis strategies for each spot. Amount and concentration of the deposited materials can be adjusted with the laser parameters. Employing similar arrays, we can analyze the human immune response towards the proteome of different pathogens. We screened several peptide array replicas with different patient sera. The screenings resulted in significant hits in several proteins with interesting implications for future diagnostics and vaccine development.

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