Cell explosion and nanophotonics for single cell metabolomics
Abstract
In my inaugural lecture, I presented the significance and technical background of single-cell metabolomics. The central idea of the research is a paradigm shift currently unfolding in biology: instead of traditional measurements that average millions of cells, the study of individual cells enables the identification of hidden phenotypes and metabolic states (subpopulations). Analyzing single cells presents a significant analytical challenge due to the extremely small volumes (fL–nL), high chemical complexity, and the lack of amplification procedures (such as PCR) in metabolomics. We have developed several breakthrough methods based on mass spectrometry (MS) to overcome these obstacles. Laser ablation electrospray ionization (LAESI) utilizes a mid-infrared laser to excite the water content of cells, enabling "explosive" sampling followed by ionization via electrospray. This process is described by a physical similarity model resembling Taylor-type point explosions. In a nanophotonics-based procedure, we employed silicon nanopost arrays (NAPA) for laser desorption ionization, which allows for extremely sensitive measurements (reaching detection limits as low as 800 zeptomoles) in single yeast or microbial cells. Through high-throughput automation, we progressed from initial slow, manual capillary microsampling (<10 cells/hour) to fully automated, image-analysis-based targeting capable of analyzing more than 800 cells per hour. To explore the "dark metabolome," measurements were performed using a 21 Tesla FT-ICR-MS instrument combined with the LAESI method. This approach utilizes isotopic fine structure (IFS) to determine the molecular formulas of unknown compounds, even from a single cell. We have successfully applied these technologies to characterize cell death processes (via adenylate energy charge or AEC) and to investigate nitrogen fixation in soybean root nodules. In the latter case, we managed to isolate the metabolic profiles of bacteria-infected and uninfected cells and map the spatial distribution of bio-oligomers (such as polyhydroxybutyrate or PHB) within the tissues. These results enable in vivo single-cell analysis and spatial metabolomics to open new horizons for systems biology and medicine.
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