We developed an end-to-end experimental pipeline allowing quantification of In vivo fluxes in photosynthesis and central carbon metabolism from cell to mass-spec. This pipeline employs cutting edge metabolomics, microfluidics and phenotyping systems and is constantly improved to meet sensitivity and throughput requirements, and to integrate with new computational tools.
CO2 labeling setups To support isotopically non-stationary metabolic flux analysis (INST-MFA), we developed a set of tailor-made metabolic-phenotyping setups based on combining gas mixers with microfluidics for liquid cultures (algae, cyanobacteria) or gas chambers for higher plants. Label is supplied via medium pre-bubbled with CO2-based synthetic air mixture (algae) or with the air mixture itself (plants).
Photosynthesis-focused Metabolomics The analytical infrastructure in the lab includes metabolomics analysis of most photosynthetic intermediates, including the CBC, sucrose and starch synthesis, photorespiration, C4 photosynthesis, and additional downstream central C metabolism pathways. Focusing mostly on polar metabolites, our main setup is a bio-compatible uHPLC system linked to a high-end tandem MS (Sciex Qtrap 6500+) equipped with Ion Mobility technology (SelexION).
A major effort is currently dedicated to implementing new separation techniques (HILIC, Anion-exchange) which will bypass the need for ion-pairing reagents involved in the separation of phosphorylated intermediates of photosynthesis, in order to extend the pool of studied metabolites and analytical applications.
Photobioreactors Our basic experimental setups for growth and physiology analyses, are state-of-the-art photobioreactors which allow full and accurate control and monitoring of algal growth over a wide range of conditions. Following the development of our microfluidic setups, we are currently designing the next generation of this system, which will allow high-throughput pulse-labeling via an automated and controlled auto-sampler. If you’re an enthusiastic experimentalist with a crave for an engineering challenge, let’s talk.