title: Quantitative Determination and Subcellular Imaging of Cu in Single Cells via Laser Ablation-ICP-Mass Spectrometry Using High-Density Microarray Gelatin Standards.
abstract: This manuscript describes the development and characterization of a high-density microarray calibration standard, manufactured in-house and designed to overcome the limitations in precision, accuracy, and throughput of current calibration approaches for the quantification of elemental concentrations on the cellular level using laser ablation-inductively coupled plasma-mass spectrometry (LA-ICPMS). As a case study, the accumulation of Cu in the model organism Scrippsiella trochoidea resulting from transition metal exposure (ranging from 0.5 to 100 μg/L) was evaluated. After the Cu exposure, cells of this photosynthetic dinoflagellate were treated with a critical point drying protocol, transferred to a carbon stub, and sputter-coated with a Au layer for scanning electron microscopy (SEM) analysis. In subsequent LA-ICPMS analysis, approximately 100 cells of each population were individually ablated. This approach permitted the evaluation of the mean concentration of Cu in the cell population across different exposure levels and also allowed the examination of the cellular distribution of Cu within the populations. In a cross-validation exercise, subcellular LA-ICPMS imaging was demonstrated to corroborate synchrotron radiation confocal X-ray fluorescence (SR-XRF) microimaging of single cells investigated under in vivo conditions.
authors.affiliation: Department of Analytical Chemistry, Ghent University , Campus Sterre, Krijgslaan 281 - S12, 9000 Ghent, Belgium.
authors.forename: Stijn J M
authors.initials: SJ
authors.lastname: Van Malderen
authors.affiliation: Department of Analytical Chemistry, Ghent University , Campus Sterre, Krijgslaan 281 - S12, 9000 Ghent, Belgium.
authors.forename: Eva
authors.initials: E
authors.lastname: Vergucht
authors.affiliation: Department of Applied Ecology and Environmental Biology, Ghent University , Jozef Plateaustraat 22, 9000 Ghent, Belgium.
authors.forename: Maarten
authors.initials: M
authors.lastname: De Rijcke
authors.affiliation: Department of Applied Ecology and Environmental Biology, Ghent University , Jozef Plateaustraat 22, 9000 Ghent, Belgium.
authors.forename: Colin
authors.initials: C
authors.lastname: Janssen
authors.affiliation: Department of Analytical Chemistry, Ghent University , Campus Sterre, Krijgslaan 281 - S12, 9000 Ghent, Belgium.
authors.forename: Laszlo
authors.initials: L
authors.lastname: Vincze
authors.affiliation: Department of Analytical Chemistry, Ghent University , Campus Sterre, Krijgslaan 281 - S12, 9000 Ghent, Belgium.
authors.forename: Frank
authors.initials: F
authors.lastname: Vanhaecke
mesh_terms: D003300:Copper / Q000032:analysis*; D004141:Dinoflagellida / Q000737:chemistry / Q000166:cytology*; D005780:Gelatin / Q000737:chemistry* / Q000592:standards*; D007834:Lasers; D013058:Mass Spectrometry / Q000379:methods* / Q000592:standards; D059010:Single-Cell Analysis / Q000379:methods* / Q000592:standards
chemical_list: D003300:Copper; D005780:Gelatin