Marjan Dolatmoradi, Graduate Student, Vertes Lab, GW Department of Chemistry
Mapping Biological Nitrogen Fixation One Cell at a Time
Metabolic heterogeneity is an inherent property of cell populations, including isogenic populations, that can be manifested in phenotypic differences. Metabolite levels determined by single-cell analysis provide information on the functioning of metabolic pathways in individual cells and can reveal much of the cellular phenomena that are masked in cell-population studies. Single-cell metabolomics based on mass spectrometry (MS) has become a powerful platform that allows for studying spatiotemporal dynamics of metabolite levels at cellular resolution. To achieve the statistical characterization of heterogeneity in metabolic cell states, numerous cells must be analyzed, and for that, high-throughput techniques are needed. In our study, we utilized optical fiber-based laser ablation electrospray ionization (f-LAESI) mass spectrometry (MS) with ion mobility separation (IMS) combined with a dual-channel microscope, capable of simultaneous brightfield and fluorescence imaging, for microscopy guided high-throughput single-cell analysis. To explore metabolic heterogeneity in biological nitrogen fixation, soybean (Glycine max) root nodule cells infected by rhizobia (Bradyrhizobium japonicum) were studied. The f-LAESI sampling method allows for direct analysis of tissue-embedded single cells in their native state, resulting in reduced artifacts due to high metabolite turnover and diffusion rates. Using high-resolution IMS, structural isomers can be separated on a millisecond timescale thereby improving the molecular coverage and confidence in metabolite identification. Population-wide heterogeneity, rare cells and hidden subpopulations can also be discovered using this technique.
Bio
Marjan Dolatmoradi received her M.S. degree in computational chemistry from American University in Washington, DC in 2018. In 2019, she joined the research group of Prof. Akos Vertes at the George Washington University. Her current work focuses on single-cell metabolomics using mass spectrometry-based techniques under ambient conditions.