Dynamic force microscopy imaging of native membranes.
We employed magnetic ACmode atomic force microscopy (MACmode AFM) as a novel dynamic force microscopy method to image surfaces of biological membranes in their native environments. The lateral resolution achieved under optimized imaging conditions was in the nanometer range, even when the sample was only weakly attached to the support. Purple membranes (PM) from Halobacterium salinarum were used as a test standard for topographical imaging. The hexagonal arrangement of the bacteriorhodopsin trimers on the cytoplasmic side of PM was resolved with 1.5nm lateral accuracy, a resolution similar to images obtained in contact and tapping-mode AFM. Human rhinovirus 2 (HRV2) particles were attached to mica surfaces via nonspecific interactions. The capsid structure and 2nm sized protein loops of HRV2 were routinely obtained without any displacement of the virus. Globular and filamentous structures on living and fixed endothelial cells were observed with a resolution of 5-20nm. These examples show that MACmode AFM is a favorable method in studying the topography of soft and weakly attached biological samples with high resolution under physiological conditions.
Kienberger
Ferry
F
Institute for Biophysics, University of Linz, Altenbergerstr. 69, A-4040, Linz, Austria.
Stroh
Cordula
C
Kada
Gerald
G
Moser
Rosita
R
Baumgartner
Werner
W
Pastushenko
Vassili
V
Rankl
Christian
C
Schmidt
Ute
U
Müller
Harald
H
Orlova
Elena
E
LeGrimellec
Christian
C
Drenckhahn
Detlev
D
Blaas
Dieter
D
Hinterdorfer
Peter
P
D002213:Capsid / Q000648:ultrastructure; D002462:Cell Membrane / Q000648:ultrastructure*; D002478:Cells, Cultured; D004727:Endothelium / Q000166:cytology; D019617:Halobacterium salinarum / Q000648:ultrastructure; D006801:Humans; D008280:Magnetics; D018625:Microscopy, Atomic Force / Q000379:methods*; D018717:Purple Membrane / Q000648:ultrastructure*; D012229:Rhinovirus / Q000648:ultrastructure