Backscattered electron imaging of cultured cells: application to electron probe X-ray microanalysis using a scanning electron microscope.
We report a simple method to study the elemental content in cultured human adherent cells by electron probe X-ray microanalysis with scanning electron microscopy. Cells were adapted to grow on polycarbonate tissue culture cell inserts, washed with distilled water, plunge-frozen with liquid nitrogen and freeze-dried. Unstained, freeze-dried cultured cells were visualized in the secondary and backscattered electron imaging modes of scanning electron microscopy. With backscattered electron imaging it was possible to identify unequivocally major subcellular compartments, i.e. the nucleus, nucleoli and cytoplasm. X-ray microanalysis was used simultaneously to determine the elemental content in cultured cells at the cellular level. In addition, we propose some improvements to optimize backscattered electron and X-ray signal collection. Our findings demonstrate that backscattered electron imaging offers a powerful method to examine whole, freeze-dried cultured cells for scanning electron probe X-ray microanalysis.
Fernández-Segura
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Department of Cell Biology, School of Medicine, University of Granada, Spain. efernandez@histolii.ugr.es
Cañizares
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Cubero
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Revelles
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Campos
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D000818:Animals; D002466:Cell Nucleolus / Q000737:chemistry* / Q000648:ultrastructure; D002467:Cell Nucleus / Q000737:chemistry* / Q000648:ultrastructure; D002478:Cells, Cultured / Q000737:chemistry / Q000648:ultrastructure; D003593:Cytoplasm / Q000737:chemistry* / Q000648:ultrastructure; D004577:Electron Probe Microanalysis / Q000379:methods*; D005612:Freeze Drying; D006801:Humans; D008855:Microscopy, Electron, Scanning; D013048:Specimen Handling / Q000379:methods*; D013552:Swine; D014407:Tumor Cells, Cultured / Q000737:chemistry / Q000648:ultrastructure