Abstract
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.
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Acknowledgments
The authors thank Dr. Andrea Cremona for his technical assistance and support given to this work and Dr. Daniel Beck from Warwick University for the revision of the manuscript. This research was funded by grants from the Italian Ministry of Research and University (FIRB programme No. RBNE03B8KK-08) to V.F.S. for the project “Investigation of protein structure and function by AFM and physiological studies.”
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Orsini, F., Santacroce, M., Arosio, P. et al. Intermittent contact mode AFM investigation of native plasma membrane of Xenopus laevis oocyte. Eur Biophys J 38, 903–910 (2009). https://doi.org/10.1007/s00249-009-0464-7
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DOI: https://doi.org/10.1007/s00249-009-0464-7