Imaging the cell surface: argon sputtering to expose inner cell structures.
Established microscopies such as Scanning Electron Microscopy (SEM) and more recent developments such as Atomic Force Microscopy (AFM) and X-ray Photo-Electron Emission spectroMicroscopy (X-PEEM) can only image the sample surface. We present an argon sputtering method able to progressively expose inner cell structures without apparent damage. By varying the sputtering time, the structure of cell cytoskeleton, vesicles, mitochondria, nuclear membrane, and nucleoli can be imaged. We compared images obtained with confocal fluorescence microscopy, transmission electron microscopy (TEM), SEM, and X-PEEM on similar samples after argon sputtering, then confirmed the similarity of reference intracellular structures, including cytoskeleton fibers, cell-cell and cell-substrate adhesion structures, and secretory vesicles. We conclude that the sputtering method is a new valuable tool for surface sensitive microscopies.
De Stasio
Gelsomina
G
University of Wisconsin-Madison, Department of Physics and Synchrotron Radiation Center, Stoughton, Wisconsin 53589, USA. pupa@src.wisc.edu
Frazer
Bradley H
BH
Girasole
Marco
M
Wiese
Lisa M
LM
Krasnowska
Ewa K
EK
Greco
Giulia
G
Serafino
Annalucia
A
Parasassi
Tiziana
T
D001128:Argon; D018938:Caco-2 Cells; D002477:Cells / Q000648:ultrastructure*; D003593:Cytoplasm / Q000648:ultrastructure; D006801:Humans; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D015388:Organelles / Q000648:ultrastructure; D014965:X-Rays
D001128:Argon