Three-dimensional mass spectral imaging of HeLa-M cells - Sample preparation, data interpretation and visualisation

John S. Fletcher, Sadia Rabbani, Alex Henderson, Nicholas P. Lockyer, John C. Vickerman

    Research output: Contribution to journalArticlepeer-review

    Abstract

    Time-of-flight secondary ion mass spectrometry (ToFSIMS) is being applied increasingly to the study of biological systems where the chemical specificity of mass spectrometry and the high lateral resolution imaging capabilities can be exploited. Here we report a comparison of two cell sample preparation methods and demonstrate how they influence the outcome of the ToFSIMS analysis for three-dimensional (3D) imaging of biological cells using our novel buncher-ToF instrument (J105 3D Chemical Imager) equipped with a C60 primary ion beam. Cells were analysed fixed and freeze-dried and non-fixed, frozen-hydrated. It is concluded that maintaining the cells in a non-fixed frozen-hydrated state during the analysis helps reduce chemical redistribution, producing cleaner spectra and improved chemical contrast in both 2D and 3D imaging. Insights into data interpretation are included and we present methods for 3D reconstruction of the data using multivariate analysis techniques. Copyright © 2011 John Wiley & Sons, Ltd.
    Original languageEnglish
    Pages (from-to)925-932
    Number of pages7
    JournalRapid communications in mass spectrometry : RCM
    Volume25
    Issue number7
    DOIs
    Publication statusPublished - Apr 2011

    Keywords

    • primary ion-beam
    • tof-sims analysis
    • spectrometry analysis
    • chemical-composition
    • single cells
    • animal-cells
    • bombardment
    • c-60
    • enhancement
    • sections

    Research Beacons, Institutes and Platforms

    • Manchester Institute of Biotechnology

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