Preparation of cell membranes for high resolution imaging by AFM

Studying the Nucleated Mammalian Cell Membrane by Single Molecule Approaches

“…Unlike the ectoplasmic side of human erythrocyte membranes [32], after digestion by PNGase F, the ectoplasmic side of MDCK cell membranes exhibited no apparent pits or indents on the smooth surface. This result indicates the absence of a dense layer of saccharides on the ectoplasmic surface of the cell membranes, consistent with the patchy distribution of carbohydrates on the membrane surface shown by STORM imaging.…”

“…As shown in Figure 1E and 1F, the ectoplasmic sides of these cell membranes were as smooth as those of MDCK cells. To test our technique, we previously decorated antibodies on the cell membranes, and the result showed that the resolution of AFM was high enough to distinguish protruding proteins from the surface of cell membranes [32]. Meanwhile, it should be noted that we imaged the native or quasi-native cell membranes (unfixed), which allowed us to detect the original state of cell membranes.…”

Changes in red blood cell membrane structure in G6PD deficiency: An atomic force microscopy study

“…Subsequently, 100 μl of the diluted RBCs was deposited onto the AP-mica for 20 min. Human RBCs attached to APTES mica were sheared open by a fast stream of 7.5 mmol/l PBS (pH 7.5) through a needle at an angle of 20°, and we obtained the cell's membrane without membrane skeleton proteins in a one-step preparation as described previously [12]. Finally, RBC membranes attached to the APTES mica were washed three times in ddH 2 O.…”

“…As a very high-resolution type of scanning probe microscopy, AFM is shown in various studies to be a powerful tool for imaging membrane proteins at nanometer lateral resolution and subangstrom Z-axis resolution [12][13][14][15]. Nanobiomechanics of cells have been identified as vital characteristics to distinguish diseased cells from normal cells because diseases can not only cause biological and functional alterations but also induce abnormalities in physical and structural characteristics of cells [16][17][18].…”

Phospholipase-Catalyzed Hydrolysis in an Artificial Cell Membrane in the Presence of Melittin

“…The study of the cell membrane structure is vital for various applications, including drug screening, cancer treatment, the analysis of the control of signal transduction in the liquid mosaic model, lipid raft modeling, and protein domain modeling, and many studies on the cell membrane structure have been constructed in the last three decades. [1][2][3][4][5] However, living cells are very sensitive to environmental changes such as changes in temperature, pH, or humidity, and this sensitivity is a major disadvantage when performing experiments. In addition, the experimental results are difficult to analyze because the cells grow and produce secondary metabolites while the experiments are run.…”