title: Single cell analysis with probe ESI-mass spectrometry: detection of metabolites at cellular and subcellular levels.
abstract: Molecular analysis at cellular and subcellular levels, whether on selected molecules or at the metabolomics scale, is still a challenge now. Here we propose a method based on probe ESI mass spectrometry (PESI-MS) for single cell analysis. Detection of metabolites at cellular and subcellular levels was successfully achieved. In our work, tungsten probes with a tip diameter of about 1 μm were directly inserted into live cells to enrich metabolites. Then the enriched metabolites were directly desorbed/ionized from the tip of the probe for mass spectrometry (MS) detection. The direct desorption/ionization of the enriched metabolites from the tip of the probe greatly improved the sensitivity by a factor of about 30 fold compared to those methods that eluted the enriched analytes into a liquid phase for subsequent MS detection. We applied the PESI-MS to the detection of metabolites in single Allium cepa cells. Different kinds of metabolites, including 6 fructans, 4 lipids, and 8 flavone derivatives in single cells, have been successfully detected. Significant metabolite diversity was observed among different cells types of A. cepa bulb and different subcellular compartments of the same cell. We found that the inner epidermal cells had about 20 fold more fructans than the outer epidermal cells, while the outer epidermal cells had more lipids. We expected that PESI-MS might be a candidate in the future studies of single cell "omics".
authors.affiliation: Beijing Key Laboratory for Microanalytical Methods and Instrumentation, Department of Chemistry, Tsinghua University , Beijing China.
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Gong
authors.forename: Yaoyao
authors.initials: Y
authors.lastname: Zhao
authors.forename: Shaoqing
authors.initials: S
authors.lastname: Cai
authors.forename: Shujie
authors.initials: S
authors.lastname: Fu
authors.forename: Chengdui
authors.initials: C
authors.lastname: Yang
authors.forename: Sichun
authors.initials: S
authors.lastname: Zhang
authors.forename: Xinrong
authors.initials: X
authors.lastname: Zhang
mesh_terms: D000804:Angiotensin II / Q000032:analysis; D002477:Cells / Q000737:chemistry*; D005630:Fructans / Q000032:analysis; D055432:Metabolomics / Q000379:methods*; D019697:Onions / Q000737:chemistry / Q000166:cytology; D018517:Plant Roots / Q000737:chemistry; D059010:Single-Cell Analysis / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013347:Subcellular Fractions / Q000737:chemistry*; D017973:Tungsten Compounds / Q000737:chemistry
chemical_list: D005630:Fructans; D017973:Tungsten Compounds; D000804:Angiotensin II