Negative mode nanostructure-initiator mass spectrometry for detection of phosphorylated metabolites

Amantonico, Andrea; Flamigni, Luca; Glaus, Reto; Zenobi, Renato

doi:10.1007/s11306-009-0163-5

Cited by 28 publications

(26 citation statements)

References 24 publications

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“…A number of analytical approaches were developed with detection limits low enough for single-cell metabolite analyses [e.g., nanostructured surfaces (7,8), postionization techniques (9,10), modified laser optics (11), the use of microsampling tools (12,13), microarrays for MS measurements (14,15), etc.]. Until now, however, most MS studies targeting single-cell metabolite analysis have only shown the analytical capabilities, but have not demonstrated that true biological information can be retrieved from studying the metabolism of single cells.…”

mentioning

confidence: 99%

Mass spectrometry-based metabolomics of single yeast cells

Ibáñez¹,

Fagerer²,

Schmidt³

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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214

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Single-cell level measurements are necessary to characterize the intrinsic biological variability in a population of cells. In this study, we demonstrate that, with the microarrays for mass spectrometry platform, we are able to observe this variability. We monitor environmentally (2-deoxy-D-glucose) and genetically (ΔPFK2) perturbed Saccharomyces cerevisiae cells at the single-cell, few-cell, and population levels. Correlation plots between metabolites from the glycolytic pathway, as well as with the observed ATP/ADP ratio as a measure of cellular energy charge, give biological insight that is not accessible from population-level metabolomic data.single-cell measurements | MALDI mass spectrometry | baker's yeast E ven genetically identical cells present in the same microenvironment can express different phenotypes, for a number of reasons: cell-to-cell heterogeneity can stem from differences in the cell age and differences in the cell cycle stage, and stochastic effects together with feedback mechanisms can lead to distinctively different phenotypes, too (1-6). As population-level measurement techniques inherently average out such cell-to-cell differences, biochemical mechanisms underlying a studied system cannot be deduced from such measurements. Thus, to detect and exploit this heterogeneity, new analytical platforms with a sensitivity at the single-cell level and the ability to perform quantitative analyses must be developed and validated.Motivated by advances of mass spectrometry (MS) in metabolomics, the analytical chemistry community has stepped up its efforts toward realizing MS-based single-cell metabolomics (1, 2). A number of analytical approaches were developed with detection limits low enough for single-cell metabolite analyses [e.g., nanostructured surfaces (7, 8), postionization techniques (9, 10), modified laser optics (11), the use of microsampling tools (12, 13), microarrays for MS measurements (14, 15), etc.]. Until now, however, most MS studies targeting single-cell metabolite analysis have only shown the analytical capabilities, but have not demonstrated that true biological information can be retrieved from studying the metabolism of single cells.Here, using the unicellular eukaryotic model organism Saccharomyces cerevisiae, we present an analytical validation of a single-cell metabolite analysis using the microarrays for mass spectrometry (MAMS) platform. This validation concerns both the analytical methodology and the biological information, by monitoring expected cellular responses upon an environmental and a genetic perturbation. Furthermore, we present examples of biological insight that are only accessible with a platform such as MAMS. Specifically, we unravel metabolite-metabolite correlations, and visualize coexisting subpopulations in an isogenic cell culture. This technology can now be used to reveal metabolic differences in cells of isogenic cell populations, such as differences caused by cell cycles stages, cell ages, or stochastically induced phenotypic differences. Results and ...

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mentioning

confidence: 99%

Mass spectrometry-based metabolomics of single yeast cells

Ibáñez¹,

Fagerer²,

Schmidt³

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

214

123

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“…This versatility is a particularly powerful characteristic of NIMS surface that expands the range of detectable metabolites and, consequently, extract the maximum information from chemical complex samples. For instance, Zenobi and co-workers [24] demonstrated that compounds containing amino groups as initiators promoted the formation of ions in negative ionization mode, which suggests that the basicity of the initiator could assist the ionization process. By using 3-aminopropyldimethylethoxysilane (APDMES) as initiator, they detected phosphorylated metabolites in the femtomole range.…”

Section: Nims Surface Modificationsmentioning

confidence: 99%

Nanostructure Initiator Mass Spectrometry for tissue imaging in metabolomics: Future prospects and perspectives

Calavia

Annanouch

Correig

et al. 2012

Journal of Proteomics

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“…However, analysis of biofluids with negative-ion mode is possible with NIMS when a compatible pre-etch cleaning method or initiator material is used 35,36. When the highly acidic “Piranha solution” (sulfuric acid/hydrogen peroxide) was replaced with methanol as the pre-etch cleaning solvent, fatty acids were readily detected from a complex biofluid with NIMS 35.…”

Section: Nims Analysis Of Complex Biofluidsmentioning

confidence: 99%

Nanostructure-Initiator Mass Spectrometry Metabolite Analysis and Imaging

2010

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Metabolites are downstream end products of gene and protein activity that closely correlate with the phenotype of a biological organism. [1][2][3][4][5][6][7] Therefore, by observing specific metabolic changes, one can gain insight into perturbations underlying disease. 8 Consequently, increasing attention has been dedicated to analyzing metabolites using MS in the context of clinical diagnostics, understanding disease mechanisms, and identifying new therapeutic targets.5 , 9 , 10 The ability to analyze metabolites directly from biofluids and tissues continues to challenge current MS technology, largely because of the limits imposed by the complexity of these samples, which contain thousands to tens of thousands of metabolites. 11 A new technology being developed to address this challenge is Nanostructure-Initiator MS (NIMS), a desorption/ionization approach that does not require the application of matrix and thereby facilitates small-molecule (i.e., metabolite) identification. 12 Surface-based mass analysis has seen a resurgence in the past decade, with new MS technologies focused on increasing sensitivity, minimizing background, and reducing sample preparation. 4,6,7,13, 14 MALDI is one of the primary MS platforms currently used for the analysis of biological samples.2 , 15 , 16 However, the application of a MALDI matrix can add significant background at <1000 Da that complicates analysis of the low-mass range (i.e., metabolites).17 In addition, the size of the resulting matrix crystals limits the spatial resolution that can be achieved in tissue imaging.14 Because of these limitations, several matrix-free desorption/ionization approaches have been applied to the analysis of biofluids and tissues.17 Secondary ion MS (SIMS) was one of the first matrix-free desorption/ ionization approaches used to analyze metabolites from biological samples.18 SIMS uses a high-energy primary ion beam to desorb and generate secondary ions from a surface. The primary advantage of SIMS is its high spatial resolution (as small as 50 nm), a powerful characteristic for tissue imaging with MS.19 However, SIMS has yet to be readily applied to the analysis of biofluids and tissues because of its limited sensitivity at >500 Da and analyte fragmentation generated by the high-energy primary ion beam.14 Desorption electrospray ionization (DESI) is a matrix-free technique for analyzing biological samples that uses a charged solvent spray to desorb ions from a surface.20 Advantages of DESI are that no special surface is required and the analysis is performed at ambient pressure with full access to the sample during acquisition.20 , 21 The main limitation of DESI is spatial resolution because "focusing" the charged solvent spray is difficult.22 However, a recent development termed laser ablation ESI (LAESI) is a promising approach to circumvent this limitation. 23 Desorption/ionization on silicon (DIOS), the precursor to NIMS, is another matrix-free laser-induced desorption/ionization approach.24 , 25 DIOS utilizes a porous silicon substrate ...

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Negative mode nanostructure-initiator mass spectrometry for detection of phosphorylated metabolites

Cited by 28 publications

References 24 publications

Mass spectrometry-based metabolomics of single yeast cells

Mass spectrometry-based metabolomics of single yeast cells

Nanostructure Initiator Mass Spectrometry for tissue imaging in metabolomics: Future prospects and perspectives

Nanostructure-Initiator Mass Spectrometry Metabolite Analysis and Imaging

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