Intermittent contact mode AFM investigation of native plasma membrane of Xenopus laevis oocyte

Intermittent contact mode atomic force microscopy (AFM) was used to visualize the native plasma membrane of Xenopus laevis oocytes. Oocyte membranes were purified via ultracentrifugation on a sucrose gradient and adsorbed on mica leaves. AFM topographs and the corresponding phase images allowed for...

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Bibliographic Details
Published in:European biophysics journal 2009-09, Vol.38 (7), p.903-910
Main Authors: Orsini, Francesco, Santacroce, M, Arosio, P, Castagna, M, Lenardi, C, Poletti, G, Sacchi, F. V
Format: Article
Language:English
Subjects:
Adsorption
Aluminum Silicates - chemistry
Animals
Biochemistry
Biological and Medical Physics
Biomedical and Life Sciences
Biophysics
Cell Biology
Cell Membrane - metabolism
Crystallography, X-Ray
Female
Life Sciences
Lipid Bilayers - chemistry
Lipid Bilayers - metabolism
Lipids
Membrane Biology
Membrane Proteins - chemistry
Membrane Proteins - metabolism
Membranes
Microscopy
Microscopy, Atomic Force
Nanotechnology
Neurobiology
Oocytes - cytology
Original Paper
Plasma
Proteins
Sucrose - chemistry
Time Factors
Ultracentrifugation
Xenopus laevis
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Summary:Intermittent contact mode atomic force microscopy (AFM) was used to visualize the native plasma membrane of Xenopus laevis oocytes. Oocyte membranes were purified via ultracentrifugation on a sucrose gradient and adsorbed on mica leaves. AFM topographs and the corresponding phase images allowed for visualization and identification of both oocyte plasma membrane patches and pure lipid bilayer regions with a height of about 5 nm within membrane patches. The quantitative analysis showed a normal distribution for the lateral dimension and height of the protein complexes centered on 16.7 ± 0.2 nm (mean ± SE, n = 263) and 5.4 ± 0.1 nm (n = 262), respectively. The phase signal, providing material-dependent information, allowed for the recognition of structural features observed in AFM topographs.
ISSN:0175-7571
1432-1017
DOI:10.1007/s00249-009-0464-7