Imaging of the membrane surface of MDCK cells by atomic force microscopy.
The membrane surface of polarized renal epithelial cells (MDCK cells) grown as a monolayer was imaged with the atomic force microscope. The surface topography of dried cells determined by this approach was consistent with electron microscopy images previously reported. Fixed and living cells in aqueous medium gave more fuzzy images, likely because of the presence of the cell glycocalix. Treatment of living cells with neuraminidase, an enzyme that partly degrades the glycocalix, allowed sub-micrometer imaging. Protruding particles, 10 to 60 nm xy size, occupy most of the membrane surface. Protease treatment markedly reduced the size of these particles, indicating that they corresponded to proteins. Tip structure effects were probably involved in the exaggerated size of imaged membrane proteins. Although further improvements in the imaging conditions, including tip sharpness, are required, atomic force microscope already offers the unique possibility to image proteins at the membrane surface of living cells.
Le Grimellec
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C
Institut National de la Santé et de la Recherche Médicale U251, Faculté de Médecine Xavier-Bichat, Paris, France.
Lesniewska
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E
Cachia
C
C
Schreiber
J P
JP
de Fornel
F
F
Goudonnet
J P
JP
D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000648:ultrastructure*; D004285:Dogs; D004848:Epithelium / Q000648:ultrastructure; D007668:Kidney; D018625:Microscopy, Atomic Force / Q000379:methods