Visualization of high resolution spatial mass spectrometric data during acquisition

Thomas, Mathew; Heath, Brandi S.; Laskin, Julia; Li, Dongsheng; Liu, Ellen; Hui, Katrina; Kuprat, Andrew P.; Dam, Kerstin Kleese van; Carson, James P.

doi:10.1109/embc.2012.6347250

Cited by 23 publications

(25 citation statements)

References 8 publications

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“…The cell containing glass slide is attached to the motorized XYZ-stage controlled using software. 45 Upon finding a target cell, the Z-stage is precisely lifted (0.1 μ m increment) for cell insertion monitored using microscopes. Cellular contents are extracted by the continuously flowing sampling solvent and drawn by the capillary action toward the nano-ESI emitter, where the extracted molecules are immediately ionized for MS analysis (Figure S1).…”

Section: Methodsmentioning

confidence: 99%

“…In particular, this reactive SCMS approach provides higher ion signal intensities for the adducted metabolites. Using a Thermo LTQ Orbitrap XL mass spectrometer and the home-built single-probe SCMS setup 35 controlled using the LabView software package, 45 high mass accuracy MS analyses were performed for tentative molecular assignments (mass error <5 ppm). MS/MS analyses were also carried out to validate assignments of species of interest.…”

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confidence: 99%

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Using Dicationic Ion-Pairing Compounds To Enhance the Single Cell Mass Spectrometry Analysis Using the Single-Probe: A Microscale Sampling and Ionization Device

et al. 2016

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A unique mass spectrometry (MS) method has been developed to determine the negatively charged species in live single cells using the positive ionization mode. The method utilizes dicationic ion-pairing compounds through the miniaturized multifunctional device, the single-probe, for reactive MS analysis of live single cells under ambient conditions. In this study, two dicationic reagents, 1,5-pentanediyl-bis(1-butylpyrrolidinium) difluoride (C5(bpyr)2F2) and 1, 3-propanediyl-bis-(tripropylphosphonium) difluoride (C3(triprp)2F2), were added in the solvent and introduced into single cells to extract cellular contents for real-time MS analysis. The negatively charged (1− charged) cell metabolites, which form stable ion-pairs (1+ charged) with dicationic compounds (2+ charged), were detected in positive ionization mode with a greatly improved sensitivity. We have tentatively assigned 192 and 70 negatively charged common metabolites as adducts with (C5(bpyr)2F2) and (C3(triprp)2F2), respectively, in three separate SCMS experiments in the positive ion mode. The total number of tentatively assigned metabolites is 285 for C5(bpyr)2F2 and 143 for C3(triprp)2F2. In addition, the selectivity of dicationic compounds in the complex formation allows for the discrimination of overlapped ion peaks with low abundances. Tandem (MS/MS) analyses at the single cell level were conducted for selected adduct ions for molecular identification. The utilization of the dicationic compounds in the single-probe MS technique provides an effective approach to the detection of a broad range of metabolites at the single cell level.

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Using Dicationic Ion-Pairing Compounds To Enhance the Single Cell Mass Spectrometry Analysis Using the Single-Probe: A Microscale Sampling and Ionization Device

et al. 2016

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“…With this flow rate, solvent delivered to the surface can be completely drawn up by the ESI-emitter with no surface spreading observed. The total time for a typical high resolution MSI experiment for a sample of 2.0 × 1.5 mm in size is around 4–5 h. MS images were generated from raw data files using the MSI QuickView software developed by PNNL [44]. Accurate mass of metabolite was searched using the online database Metlin from the Scripps Center for Metabolomics (https://metlin.scripps.edu/index.php).…”

Section: Methodsmentioning

confidence: 99%

High-Resolution Ambient MS Imaging of Negative Ions in Positive Ion Mode: Using Dicationic Reagents with the Single-Probe

Rao

Pan

Tian

et al. 2015

J. Am. Soc. Mass Spectrom.

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We have used the Single-probe, a miniaturized sampling device utilizing in-situ surface microextraction for ambient mass spectrometry (MS) analysis, for the high resolution MS imaging (MSI) of negatively charged species in the positive ionization mode. Two dicationic compounds, 1,5-pentanediyl-bis(1-butylpyrrolidinium) difluoride [C5(bpyr)2F2] and 1,3-propanediyl-bis(tripropylphosphonium) difluoride [C3(triprp)2F2], were added into the sampling solvent to form 1+ charged adducts with the negatively charged species extracted from tissues. We were able to detect 526 and 322 negatively charged species this way using [C5(bpyr)2F2] and [C3(triprp)2F2], respectively, including oleic acid, arachidonic acid, and several species of phosphatidic acid, phosphoethanolamine, phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, and others. In conjunction with the identification of the non-adduct cations, we have tentatively identified a total number of 1200 and 828 metabolites from mouse brain sections using [C5(bpyr)2F2] and [C3(triprp)2F2], respectively, through high mass accuracy measurements (mass error <5 ppm); MS/MS analyses were also performed to verify the identity of selected species. In addition to the high mass accuracy measurement, we were able to generate high spatial resolution (~17 µm) MS images of mouse brain sections. Our study demonstrated that utilization of dicationic compounds in the surface microextraction with the Single-probe device can perform high mass and spatial resolution ambient MSI measurements of broader types of compounds in tissues. Other reagents can be potentially used with the Single-probe device for a variety of reactive MSI studies to enable the analysis of species that are previously intractable.

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“…The solvent used was methanol:water (9:1) supplied at a flow rate of 0.2 μL/min. After data acquisition, the raw MS files were converted to MS images using the MSI QuickView software developed by PNNL [28].…”

Section: Ms Imaging and Data Interpretationmentioning

confidence: 99%

High Resolution Tissue Imaging Using the Single-probe Mass Spectrometry under Ambient Conditions

Rao

Pan

Yang

2015

J. Am. Soc. Mass Spectrom.

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Abstract. Ambient mass spectrometry imaging (MSI) is an emerging field with great potential for the detailed spatial analysis of biological samples with minimal pretreatment. We have developed a miniaturized sampling and ionization device, the Singleprobe, which uses in-situ surface micro-extraction to achieve high detection sensitivity and spatial resolution during MSI experiments. The Single-probe was coupled to a Thermo LTQ Orbitrap XL mass spectrometer and was able to create high spatial and high mass resolution MS images at 8±2 and 8.5 μm on flat polycarbonate microscope slides and mouse kidney sections, respectively, which are among the highest resolutions available for ambient MSI techniques. Our proof-of-principle experiments indicate that the Single-probe MSI technique has the potential to obtain ambient MS images with very high spatial resolutions with minimal sample preparation, which opens the possibility for subcellular ambient tissue MSI to be performed in the future.

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Visualization of high resolution spatial mass spectrometric data during acquisition

Cited by 23 publications

References 8 publications

Using Dicationic Ion-Pairing Compounds To Enhance the Single Cell Mass Spectrometry Analysis Using the Single-Probe: A Microscale Sampling and Ionization Device

Using Dicationic Ion-Pairing Compounds To Enhance the Single Cell Mass Spectrometry Analysis Using the Single-Probe: A Microscale Sampling and Ionization Device

High-Resolution Ambient MS Imaging of Negative Ions in Positive Ion Mode: Using Dicationic Reagents with the Single-Probe

High Resolution Tissue Imaging Using the Single-probe Mass Spectrometry under Ambient Conditions

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