title: Single-Cell Analysis: Visualizing Pharmaceutical and Metabolite Uptake in Cells with Label-Free 3D Mass Spectrometry Imaging.
abstract: Detecting metabolites and parent compound within a cell type is now a priority for pharmaceutical development. In this context, three-dimensional secondary ion mass spectrometry (SIMS) imaging was used to investigate the cellular uptake of the antiarrhythmic agent amiodarone, a phospholipidosis-inducing pharmaceutical compound. The high lateral resolution and 3D imaging capabilities of SIMS combined with the multiplex capabilities of ToF mass spectrometric detection allows for the visualization of pharmaceutical compound and metabolites in single cells. The intact, unlabeled drug compound was successfully detected at therapeutic dosages in macrophages (cell line: NR8383). Chemical information from endogenous biomolecules was used to correlate drug distributions with morphological features. From this spatial analysis, amiodarone was detected throughout the cell, with the majority of the compound found in the membrane and subsurface regions and absent in the nuclear regions. Similar results were obtained when the macrophages were doped with amiodarone metabolite, desethylamiodarone. The fwhm lateral resolution measured across an intracellular interface in high lateral resolution ion images was approximately 550 nm. Overall, this approach provides the basis for studying cellular uptake of pharmaceutical compounds and their metabolites on the single cell level.
authors.affiliation: †National Centre of Excellence in Mass Spectrometry Imaging (NiCE-MSI), National Physical Laboratory (NPL), Teddington, Middlesex, TW11 0LW, United Kingdom.
authors.forename: Melissa K
authors.initials: MK
authors.lastname: Passarelli
authors.affiliation: ‡GlaxoSmithKline, Stevenage, United Kingdom.
authors.forename: Carla F
authors.initials: CF
authors.lastname: Newman
authors.affiliation: ‡GlaxoSmithKline, Stevenage, United Kingdom.
authors.forename: Peter S
authors.initials: PS
authors.lastname: Marshall
authors.affiliation: ‡GlaxoSmithKline, Stevenage, United Kingdom.
authors.forename: Andrew
authors.initials: A
authors.lastname: West
authors.affiliation: †National Centre of Excellence in Mass Spectrometry Imaging (NiCE-MSI), National Physical Laboratory (NPL), Teddington, Middlesex, TW11 0LW, United Kingdom.|§University of Nottingham, School of Pharmacy University Park, Nottingham, NG7 2RD, United Kingdom.
authors.forename: Ian S
authors.initials: IS
authors.lastname: Gilmore
authors.affiliation: †National Centre of Excellence in Mass Spectrometry Imaging (NiCE-MSI), National Physical Laboratory (NPL), Teddington, Middlesex, TW11 0LW, United Kingdom.|§University of Nottingham, School of Pharmacy University Park, Nottingham, NG7 2RD, United Kingdom.
authors.forename: Josephine
authors.initials: J
authors.lastname: Bunch
authors.affiliation: §University of Nottingham, School of Pharmacy University Park, Nottingham, NG7 2RD, United Kingdom.
authors.forename: Morgan R
authors.initials: MR
authors.lastname: Alexander
authors.affiliation: ‡GlaxoSmithKline, Stevenage, United Kingdom.
authors.forename: Colin T
authors.initials: CT
authors.lastname: Dollery
mesh_terms: D000818:Animals; D002461:Cell Line, Transformed; D013058:Mass Spectrometry / Q000379:methods*; D010599:Pharmacokinetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D059010:Single-Cell Analysis