Single-Cell Analysis: Visualizing Pharmaceutical and Metabolite Uptake in Cells with Label-Free 3D Mass Spectrometry Imaging

Passarelli, Melissa K.; Newman, Carla F.; Marshall, Peter S.; West, Andrew; Gilmore, Ian S.; Bunch, Josephine; Alexander, Morgan R.; Dollery, C. T.

doi:10.1021/acs.analchem.5b00842

Cited by 137 publications

(119 citation statements)

References 39 publications

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“…50 The ability of ToF-SIMS to detect compounds in bacteria, the approximate size of mitochondria, represents a significant technological advance beyond applying ToF-SIMS imaging to mammalian cells. 51,52 The development of low-damage gas cluster ion beams and further enhancement in ionization capability could expand the opportunities for this technique as a valuable analytical tool for biological and pharmaceutical sciences.…”

Section: Discussionmentioning

confidence: 99%

Subcellular Chemical Imaging of Antibiotics in Single Bacteria Using C₆₀-Secondary Ion Mass Spectrometry

et al. 2017

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The inherent difficulty of discovering new and effective antibacterials and the rapid development of resistance particularly in Gram-negative bacteria, illustrates the urgent need for new methods that enable rational drug design. Here we report the development of 3D imaging cluster Time-of-Flight secondary ion mass spectrometry (ToF-SIMS) as a label-free approach to chemically map small molecules in aggregated and single Escherichia coli cells, with ∼300 nm spatial resolution and high chemical sensitivity. The feasibility of quantitative analysis was explored, and a nonlinear relationship between treatment dose and signal for tetracycline and ampicillin, two clinically used antibacterials, was observed. The methodology was further validated by the observation of reduction in tetracycline accumulation in an E. coli strain expressing the tetracycline-specific efflux pump (TetA) compared to the isogenic control. This study serves as a proof-of-concept for a new strategy for chemical imaging at the nanoscale and has the potential to aid discovery of new antibacterials.

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Section: Discussionmentioning

confidence: 99%

Subcellular Chemical Imaging of Antibiotics in Single Bacteria Using C₆₀-Secondary Ion Mass Spectrometry

et al. 2017

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“…31-34 MALDI uses focused laser light to desorb and ionize sample analytes incorporated into a suitable matrix. In contrast, SIMS utilizes a beam of accelerated primary ions or larger clusters that bombards the sample surface to sputter and generate secondary ions in the gas phase for mass-to-charge ( m/z ) analysis.…”

Section: Introductionmentioning

confidence: 99%

“…32,33 The approach circumvents the need for MS raster imaging 42,43 of a large region of interest, which is time consuming and often splits cell signals over multiple pixels. However, most single cell SIMS studies also utilize raster imaging 22,34,35,44-50 to fully leverage the subcellular spatial resolution of the method and localize analytes within single cells, albeit at low throughput and reduced sensitivity.…”

Section: Introductionmentioning

confidence: 99%

Single Cell Profiling Using Ionic Liquid Matrix-Enhanced Secondary Ion Mass Spectrometry for Neuronal Cell Type Differentiation

et al. 2017

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A high-throughput single cell profiling method has been developed for matrix-enhanced secondary ion mass spectrometry (ME-SIMS) to investigate the lipid profiles of neuronal cells. Populations of cells are dispersed onto the substrate, their locations determined using optical microscopy, and the cell locations used to guide the acquisition of SIMS spectra from the cells. Up to 2,000 cells can be assayed in one experiment at a rate of 6 s per cell. Multiple saturated and unsaturated phosphatidylcholines (PCs) and their fragments are detected and verified with tandem mass spectrometry from individual cells when ionic liquids are employed as a matrix. Optically guided single cell profiling with ME-SIMS is suitable for a range of cell sizes, from Aplysia californica neurons larger than 75 μm to 7-μm rat cerebellar neurons. ME-SIMS analysis followed by t-distributed stochastic neighbor embedding of peaks in the lipid molecular mass range (m/z 700–850) distinguishes several cell types from the rat central nervous system, largely based on the relative proportions of the four dominant lipids, PC(32:0), PC(34:1), PC(36:1), and PC(38:5). Furthermore, subpopulations within each cell type are tentatively classified consistent with their endogenous lipid ratios. The results illustrate the efficacy of a new approach to classify single cell populations and subpopulations using SIMS profiling of lipid and metabolite contents. These methods are broadly applicable for high throughput single cell chemical analyses.

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“…Gostek et al showed that single bladder cancer cells can be distinguished using TOF-SIMS data and principal component analysis [50]. A recent 3D-MSI-TOF-SIMS study by Passarelli et al using dual beam analysis and depth profiling described drug and metabolite uptake at the single cell level [51]. …”

Section: Introductionmentioning

confidence: 99%

Analysis of Chemotherapeutic Drug Delivery at the Single Cell Level Using 3D-MSI-TOF-SIMS

Vanbellingen

Castellanos

Rodriguez-Silva

et al. 2016

J. Am. Soc. Mass Spectrom.

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In the present work, we show the advantages of label-free, tridimensional mass spectrometry imaging using dual beam analysis (25 keV Bi3+) and depth profiling (20 keV with a distribution centered at Ar1500+) coupled to time of flight secondary ion mass spectrometry (3D-MSI-TOF-SIMS) for the study of A-172 human glioblastoma cell line treated with B-cell lymphoma 2 (Bcl-2) inhibitor ABT-737. The high spatial (~250 nm) and high mass resolution (m/Δm ~ 10,000) of TOF-SIMS permitted the localization and identification of the intact, unlabeled drug molecular ion (m/z 811.26 C42H44ClN6O5S2−[M-H]−) as well as characteristic fragment ions. We propose a novel approach based on the inspection of the drug secondary ion yield which showed a good correlation with the drug concentration during cell treatment at therapeutic dosages (0 – 200 μM with 4 h incubation). Chemical maps using endogenous molecular markers showed that the ABT-737 is mainly localized in subsurface regions and absent in the nucleus. A semi-quantitative workflow is proposed to account for the biological cell diversity based on the spatial distribution of endogenous molecular markers (e.g., nuclei and cytoplasm) and secondary ion confirmation based on the ratio of drug-specific fragments to molecular ion as a function of the therapeutic dosage.

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Single-Cell Analysis: Visualizing Pharmaceutical and Metabolite Uptake in Cells with Label-Free 3D Mass Spectrometry Imaging

Cited by 137 publications

References 39 publications

Subcellular Chemical Imaging of Antibiotics in Single Bacteria Using C₆₀-Secondary Ion Mass Spectrometry

Subcellular Chemical Imaging of Antibiotics in Single Bacteria Using C₆₀-Secondary Ion Mass Spectrometry

Single Cell Profiling Using Ionic Liquid Matrix-Enhanced Secondary Ion Mass Spectrometry for Neuronal Cell Type Differentiation

Analysis of Chemotherapeutic Drug Delivery at the Single Cell Level Using 3D-MSI-TOF-SIMS

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Single-Cell Analysis: Visualizing Pharmaceutical and Metabolite Uptake in Cells with Label-Free 3D Mass Spectrometry Imaging

Cited by 137 publications

References 39 publications

Subcellular Chemical Imaging of Antibiotics in Single Bacteria Using C60-Secondary Ion Mass Spectrometry

Subcellular Chemical Imaging of Antibiotics in Single Bacteria Using C60-Secondary Ion Mass Spectrometry

Single Cell Profiling Using Ionic Liquid Matrix-Enhanced Secondary Ion Mass Spectrometry for Neuronal Cell Type Differentiation

Analysis of Chemotherapeutic Drug Delivery at the Single Cell Level Using 3D-MSI-TOF-SIMS

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Subcellular Chemical Imaging of Antibiotics in Single Bacteria Using C₆₀-Secondary Ion Mass Spectrometry

Subcellular Chemical Imaging of Antibiotics in Single Bacteria Using C₆₀-Secondary Ion Mass Spectrometry