Imaging of the surface of living cells by low-force contact-mode atomic force microscopy.
The membrane surface of living CV-1 kidney cells in culture was imaged by contact-mode atomic force microscopy using scanning forces in the piconewton range. A simple procedure was developed for imaging of the cell surface with forces as low as 20-50 pN, i.e., two orders of magnitude below those commonly used for cell imaging. Under these conditions, the indentation of the cells by the tip could be reduced to less than l0 nm, even at the cell center, which gave access to the topographic image of the cell surface. This surface appeared heterogeneous with very few villosities and revealed, only in distinct areas, the submembrane cytoskeleton. At intermediate magnifications, corresponding to 20-5 microm scan sizes, the surface topography likely reflected the organization of submembrane and intracellular structures on which the plasma membrane lay. By decreasing the scan size, a lateral resolution better than 20 nm was routinely obtained for the cell surface, and a lateral resolution better than 10 nm was obtained occasionally. The cell surface appeared granular, with packed particles, likely corresponding to proteins or protein-lipid complexes, between approximately 5 and 30 nm xy size.
Le Grimellec
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C
Centre de Biochimie Structurale, INSERM U414, I.U.R.C., 34093 Montpellier Cedex 05, France.
Lesniewska
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E
Giocondi
M C
MC
Finot
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E
Vié
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V
Goudonnet
J P
JP
D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000648:ultrastructure*; D002522:Chlorocebus aethiops; D007668:Kidney; D018625:Microscopy, Atomic Force / Q000379:methods*; D012680:Sensitivity and Specificity