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Title: Development and refinement of an in situ ‘molecular microscope’ utilizing ultrahigh resolution mass spectrometry

Abstract

Development and refinement of an in situ ‘molecular microscope’ utilizing ultrahigh resolution mass spectrometry Professor Gary Stacey, Principal Investigator, University of Missouri, Columbia Professor Akos Vertes, Co-Principal Investigator, George Washington University Ljiljana Paša-Tolić, Ph.D.; Christopher Anderton, Ph.D.; David W. Koppenaal, Ph.D., Co-Principal Investigators, Environmental and Molecular Science Laboratory, Pacific Northwest National Laboratory The ability to elucidate complex metabolic behavior of living organisms in situ and with cellular resolution is a bioanalytical grand challenge that is yet to be realized. Mass spectrometry imaging (MSI) using laser ablation electrospray ionization mass spectrometry (LAESI) has recently been demonstrated, and holds promise for direct spatially resolved molecular profiling in a variety of living systems of DOE relevance (e.g., plant leaf, stem, and root tissue).The LAESI MSI approach has revealed cell-to-cell heterogeneity of metabolites within living plant tissue, in a multiplexed fashion and without the need for labeling. Cellular resolution by in situ, ambient pressure MSI methods are far from routine, however, and have inadequate spatial resolution for many biological questions of interest. We propose to develop next-generation LAESI approaches, using advanced fiber optics and transmission-based geometries, in efforts to attain dynamic bioanalysis of living systems at better than 20 µm lateral resolution. In parallelmore » with LAESI innovations, attendant increases in sensitivity and dynamic range of the mass spectrometer system will also be necessary for detection of the limited amount of material being ionized. To respond to this need, a new, high magnetic field (21 Tesla) Fourier Transform Ion Cyclotron Resonance (FTICR) MS at the EMSL/PNNL will be coupled with the newly developed LAESI source. This mass spectrometer, currently being assembled, will be one of only two such systems in the world, and the only one in a DOE laboratory or user facility. Fundamentally, all key figures of merits of FTICR MS performance improve with increased magnetic field strength, providing the ability to achieve unparalleled molecular specificity (near unequivocal molecular constituent identification) at the highest possible mass resolving power and sensitivity. The work proposed here will greatly increase the utility of this FTICR MS instrument, providing unique capabilities to support BER-relevant science. The research plan is comprised of three tasks, two of which are directed by the academic leads (University of Missouri and George Washington University), and one of which will be performed in conjunction with the BER user facility at EMSL/PNNL. In task 1, advancements of the LAESI technology will be achieved, with oversight and leadership of Professor Akos Vertes of George Washington University. In task 2, incorporation of this ion source with EMSL’s new 21T FTICR MS system will be undertaken, supplemented by global omics measurements and calibrated against more conventional MSI platforms also available at EMSL (Drs. Pasa-Tolic, Anderton, and Koppenaal team). In the final task, the resultant innovations and advancements will be tested and validated by addressing important BER relevant science questions on model systems of interest under the direction of Professor Gary Stacey of the University of Missouri. This research follows on key recommendations of the recent BER “Research in Sustainable Bioenergy” report that, for example, stressed that “The design of sustainable biofuel systems requires knowledge about key plant-microbe-environment interactions that provide a range of ecosystem services.” This task will focus on validation of the instrumentation developed through this proposal, advertising this capability to the community through demonstration and, finally, discovering new biology relevant to the BER mission. This final objective will make use of Setaria viridis, a model C4 grass species, as well as switchgrass and sorghum, two promising biofuel crops. Specifically, we will demonstrate applicability to plant-microbe rhizosphere interactions focusing on the biological nitrogen fixation by monitoring metabolic changes in real-time with nutrient, growth condition, and other perturbations.« less

Authors:
ORCiD logo [1];  [2];  [3];  [3];  [3]
  1. University of Missouri
  2. George Washington Univ., Washington, DC (United States)
  3. Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Environmental Molecular Sciences Lab. (EMSL)
Publication Date:
Research Org.:
Univ. of Missouri, Columbia, MO (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23). Biological Systems Science Division
OSTI Identifier:
1532356
Report Number(s):
DOE-MU-13978
DOE Contract Number:  
SC0013978
Resource Type:
Technical Report
Resource Relation:
Related Information: Sylwia A. Stopka, Beverly J. Agtuca, David W. Koppenaal, Ljiljana Paša-Tolić, Gary Stacey, Akos Vertes, and Christopher R. Anderton. (2017) Laser ablation electrospray ionization mass spectrometry with ion mobility separation reveals metabolites and lipids in the symbiotic interactions of soybean roots and rhizobia. Plant J. 91: 340-354. DOI: 10.1111/tpj.13569 S. A. Stopka, L. Samarah, J. B. Shaw, A. Liyu, D. Veličković, B. J. Agtuca, D. Koppenaal, G. Stacey, L. Paša-Tolić, C. R. Anderton, A. Vertes. "Ambient Metabolic Profiling and Imaging of Biological Samples with Ultrahigh Molecular Resolution using Laser Ablation Electrospray Ionization 21T-FTICR Mass Spectrometry," Anal Chem 2019, 91, 5028. https://doi.org/10.1021/acs.analchem.8b05084 D. Veličković, B. Agtuca, S. A. Stopka, A. Vertes, D. W. Koppenaal, L. Paša-Tolić, G. Stacey, C. R. Anderton. “Visualization of soybean nodule metabolism by high-resolution molecular tomography,” ISME 2018, 12, 233. https://doi.org/10.1038/s41396-018-0188-8 J. A. Fincher, A. R. Korte, B. Reschke, N. J. Morris, M. J. Powell, A. Vertes. “Enhanced Sensitivity and Metabolite Coverage with Remote Laser Ablation Electrospray Ionization-Mass Spectrometry Aided by Coaxial Plume and Gas Dynamics,” Analyst, 2017, 142, 3157. DOI: 10.1039/C7AN00805H
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; 47 OTHER INSTRUMENTATION; 54 ENVIRONMENTAL SCIENCES; 59 BASIC BIOLOGICAL SCIENCES; mass spectrometry, plant, biological nitrogen fixation, soybean, symbiosis, imaging

Citation Formats

Stacey, Gary, Vertes, Akos, Paša-Tolić, Ljiljana, Anderton, Christopher, and Koppenaal, David. Development and refinement of an in situ ‘molecular microscope’ utilizing ultrahigh resolution mass spectrometry. United States: N. p., 2019. Web. doi:10.2172/1532356.
Stacey, Gary, Vertes, Akos, Paša-Tolić, Ljiljana, Anderton, Christopher, & Koppenaal, David. Development and refinement of an in situ ‘molecular microscope’ utilizing ultrahigh resolution mass spectrometry. United States. doi:10.2172/1532356.
Stacey, Gary, Vertes, Akos, Paša-Tolić, Ljiljana, Anderton, Christopher, and Koppenaal, David. Sun . "Development and refinement of an in situ ‘molecular microscope’ utilizing ultrahigh resolution mass spectrometry". United States. doi:10.2172/1532356. https://www.osti.gov/servlets/purl/1532356.
@article{osti_1532356,
title = {Development and refinement of an in situ ‘molecular microscope’ utilizing ultrahigh resolution mass spectrometry},
author = {Stacey, Gary and Vertes, Akos and Paša-Tolić, Ljiljana and Anderton, Christopher and Koppenaal, David},
abstractNote = {Development and refinement of an in situ ‘molecular microscope’ utilizing ultrahigh resolution mass spectrometry Professor Gary Stacey, Principal Investigator, University of Missouri, Columbia Professor Akos Vertes, Co-Principal Investigator, George Washington University Ljiljana Paša-Tolić, Ph.D.; Christopher Anderton, Ph.D.; David W. Koppenaal, Ph.D., Co-Principal Investigators, Environmental and Molecular Science Laboratory, Pacific Northwest National Laboratory The ability to elucidate complex metabolic behavior of living organisms in situ and with cellular resolution is a bioanalytical grand challenge that is yet to be realized. Mass spectrometry imaging (MSI) using laser ablation electrospray ionization mass spectrometry (LAESI) has recently been demonstrated, and holds promise for direct spatially resolved molecular profiling in a variety of living systems of DOE relevance (e.g., plant leaf, stem, and root tissue).The LAESI MSI approach has revealed cell-to-cell heterogeneity of metabolites within living plant tissue, in a multiplexed fashion and without the need for labeling. Cellular resolution by in situ, ambient pressure MSI methods are far from routine, however, and have inadequate spatial resolution for many biological questions of interest. We propose to develop next-generation LAESI approaches, using advanced fiber optics and transmission-based geometries, in efforts to attain dynamic bioanalysis of living systems at better than 20 µm lateral resolution. In parallel with LAESI innovations, attendant increases in sensitivity and dynamic range of the mass spectrometer system will also be necessary for detection of the limited amount of material being ionized. To respond to this need, a new, high magnetic field (21 Tesla) Fourier Transform Ion Cyclotron Resonance (FTICR) MS at the EMSL/PNNL will be coupled with the newly developed LAESI source. This mass spectrometer, currently being assembled, will be one of only two such systems in the world, and the only one in a DOE laboratory or user facility. Fundamentally, all key figures of merits of FTICR MS performance improve with increased magnetic field strength, providing the ability to achieve unparalleled molecular specificity (near unequivocal molecular constituent identification) at the highest possible mass resolving power and sensitivity. The work proposed here will greatly increase the utility of this FTICR MS instrument, providing unique capabilities to support BER-relevant science. The research plan is comprised of three tasks, two of which are directed by the academic leads (University of Missouri and George Washington University), and one of which will be performed in conjunction with the BER user facility at EMSL/PNNL. In task 1, advancements of the LAESI technology will be achieved, with oversight and leadership of Professor Akos Vertes of George Washington University. In task 2, incorporation of this ion source with EMSL’s new 21T FTICR MS system will be undertaken, supplemented by global omics measurements and calibrated against more conventional MSI platforms also available at EMSL (Drs. Pasa-Tolic, Anderton, and Koppenaal team). In the final task, the resultant innovations and advancements will be tested and validated by addressing important BER relevant science questions on model systems of interest under the direction of Professor Gary Stacey of the University of Missouri. This research follows on key recommendations of the recent BER “Research in Sustainable Bioenergy” report that, for example, stressed that “The design of sustainable biofuel systems requires knowledge about key plant-microbe-environment interactions that provide a range of ecosystem services.” This task will focus on validation of the instrumentation developed through this proposal, advertising this capability to the community through demonstration and, finally, discovering new biology relevant to the BER mission. This final objective will make use of Setaria viridis, a model C4 grass species, as well as switchgrass and sorghum, two promising biofuel crops. Specifically, we will demonstrate applicability to plant-microbe rhizosphere interactions focusing on the biological nitrogen fixation by monitoring metabolic changes in real-time with nutrient, growth condition, and other perturbations.},
doi = {10.2172/1532356},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2019},
month = {6}
}

Works referenced in this record: