Atomic force microscopy imaging ofXenopus laevis oocyte plasma membrane purified by ultracentrifugation
| Autor: | S. A. Mari, M. Marinone, Francesco Orsini, Cristina Lenardi, G. Poletti, Sara Bettè, V. F. Sacchi, M. Santacroce |
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| Rok vydání: | 2008 |
| Předmět: |
Histology
Macromolecular Substances Lipid Bilayers Xenopus Analytical chemistry Microscopy Atomic Force Xenopus laevis Physisorption Centrifugation Density Gradient Animals Sample preparation Square Micrometer Instrumentation biology Chemistry Bilayer Cell Membrane biology.organism_classification Medical Laboratory Technology Crystallography Membrane Oocytes Female Ultracentrifuge Mica Anatomy |
| Zdroj: | Microscopy Research and Technique. 71:397-402 |
| ISSN: | 1097-0029 1059-910X |
| DOI: | 10.1002/jemt.20559 |
| Popis: | Atomic force microscopy (AFM) was used to investigate the native plasma membrane of Xenopus laevis (X. laevis) oocyte purified by means of ultracentrifugation on sucrose gradient and subsequently adsorbed on mica leaves through a physisorption process. Reproducible AFM topography images were collected, analyzed, and compared. AFM images showed the presence of large single or double bilayer membrane sheets covered with protein complexes. The lateral dimension and height of protein complexes imaged in air showed a normal distribution centred on 15.4 ± 0.4 nm (mean ± SE; n = 59) and 3.9 ± 0.2 nm (mean ± SE; n = 57), respectively. A density of about 270 protein complexes per square micron was calculated. Less frequently, ordered nanometer domains with densely packed protein complexes arranged in hexagonal patterns were also visualized in AFM images, confirming previously published data. Their lateral dimension and height showed a normal distribution centred on 23.0 ± 0.4 nm (mean ± SE; n = 42) and 1.5 ± 0.6 nm (mean ± SE; n = 90), respectively. A density of about 870 protein complexes per square micrometer was calculated. Advantages and drawbacks of this new sample preparation for AFM imaging are discussed. Microsc. Res. Tech., 2008. © 2008 Wiley-Liss, Inc. |
| Databáze: | OpenAIRE |
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