Compositional mapping of the surface and interior of mammalian cells at submicrometer resolution.
We present progress toward imaging of chemical species within intact mammalian cells using secondary ion mass spectrometry, including the simultaneous mapping of subcellular elemental and molecular species along with intrinsic membrane-specific cellular markers. Results from imaging both the cell surface and cell interior exposed by site-specific focused ion beam milling demonstrate that in-plane resolutions of approximately 400-500 nm can be achieved. The results from mapping cell surface phosphatidylcholine and several other molecular ions present in the cells establish that spatially resolved chemical signatures of individual cells can be derived from novel multivariate analysis and classification of the molecular images obtained at different m/z ratios. The methods we present here for specimen preparation and chemical imaging of cell interiors provide the foundation for obtaining 3D molecular maps of unstained mammalian cells, with particular relevance for probing the subcellular distributions of small molecules, such as drugs and metabolites.
Surface and Microanalysis Science Division, National Institute of Standards and Technology, Gaithersburg, Maryland 20899-8371, USA. cszakal@nist.gov
Christopher
C
Szakal
Kedar
K
Narayan
Jing
J
Fu
Jonathan
J
Lefman
Sriram
S
Subramaniam
D000465:Algorithms; D001692:Biological Transport; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D042541:Intracellular Space / Q000737:chemistry / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods*; D057054:Molecular Imaging / Q000379:methods*; D059010:Single-Cell Analysis
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