title: Single-Cell Mass Spectrometry of Metabolites Extracted from Live Cells by Fluidic Force Microscopy.
abstract: Single-cell metabolite analysis provides valuable information on cellular function and response to external stimuli. While recent advances in mass spectrometry reached the sensitivity required to investigate metabolites in single cells, current methods commonly isolate and sacrifice cells, inflicting a perturbed state and preventing complementary analyses. Here, we propose a two-step approach that combines nondestructive and quantitative withdrawal of intracellular fluid with subpicoliter resolution using fluidic force microscopy, followed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The developed method enabled the detection and identification of 20 metabolites recovered from the cytoplasm of individual HeLa cells. The approach was further validated in13C-glucose feeding experiments, which showed incorporation of labeled carbon atoms into different metabolites. Metabolite sampling, followed by mass spectrometry measurements, enabled the preservation of the physiological context and the viability of the analyzed cell, providing opportunities for complementary analyses of the cell before, during, and after metabolite analysis.
authors.affiliation: Department of Biology, Institute of Microbiology, ETH Zurich , Vladimir-Prelog-Weg 1-5/10, 8093 Zurich, Switzerland.
authors.forename: Orane
authors.initials: O
authors.lastname: Guillaume-Gentil
authors.affiliation: Department of Biology, Institute of Microbiology, ETH Zurich , Vladimir-Prelog-Weg 1-5/10, 8093 Zurich, Switzerland.
authors.forename: Timo
authors.initials: T
authors.lastname: Rey
authors.affiliation: Department of Biology, Institute of Microbiology, ETH Zurich , Vladimir-Prelog-Weg 1-5/10, 8093 Zurich, Switzerland.
authors.forename: Patrick
authors.initials: P
authors.lastname: Kiefer
authors.affiliation: Department of Chemistry and Applied Biosciences, Laboratory of Organic Chemistry, ETH Zurich , 8093 Zurich, Switzerland.
authors.forename: Alfredo J
authors.initials: AJ
authors.lastname: Ibáñez
authors.affiliation: Department of Chemistry and Applied Biosciences, Laboratory of Organic Chemistry, ETH Zurich , 8093 Zurich, Switzerland.
authors.forename: Robert
authors.initials: R
authors.lastname: Steinhoff
authors.affiliation: Swiss Federal Laboratories for Material Science and Technology EMPA , 8600 Dübendorf, Switzerland.
authors.forename: Rolf
authors.initials: R
authors.lastname: Brönnimann
authors.affiliation: Department of Information Technology and Electrical Engineering, Institute for Biomedical Engineering, ETH Zurich , 8093 Zurich, Switzerland.
authors.forename: Livie
authors.initials: L
authors.lastname: Dorwling-Carter
authors.affiliation: Department of Information Technology and Electrical Engineering, Institute for Biomedical Engineering, ETH Zurich , 8093 Zurich, Switzerland.
authors.forename: Tomaso
authors.initials: T
authors.lastname: Zambelli
authors.affiliation: Department of Chemistry and Applied Biosciences, Laboratory of Organic Chemistry, ETH Zurich , 8093 Zurich, Switzerland.
authors.forename: Renato
authors.identifier: 0000-0001-5211-4358
authors.initials: R
authors.lastname: Zenobi
authors.affiliation: Department of Biology, Institute of Microbiology, ETH Zurich , Vladimir-Prelog-Weg 1-5/10, 8093 Zurich, Switzerland.
authors.forename: Julia A
authors.identifier: 0000-0002-6011-4910
authors.initials: JA
authors.lastname: Vorholt
mesh_terms: D002247:Carbon Isotopes; D006367:HeLa Cells; D006801:Humans; D055442:Metabolome; D055432:Metabolomics / Q000379:methods*; D008853:Microscopy / Q000379:methods*; D059010:Single-Cell Analysis / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D002247:Carbon Isotopes; C000615229:Carbon-13