scholarly journals Creating supported plasma membrane bilayers using acoustic pressure

2020 ◽  
Author(s):  
Erdinc Sezgin ◽  
Dario Carugo ◽  
Ilya Levental ◽  
Eleanor Stride ◽  
Christian Eggeling

AbstractModel membrane systems are essential tools for biology, enabling study of biological processes in a simplified setting to reveal the underlying physicochemical principles. As cell-derived membrane systems, giant plasma membrane vesicles (GPMVs) constitute an intermediate model between native cellular plasma and artificial membranes. Certain applications, however, require planar membrane surfaces. Here, we report a novel approach for creating supported plasma membrane bilayers (SPMBs) by bursting cell-derived GPMVs using an ultrasonic pressure field generated within an acoustofluidic device. We show that the mobility of outer leaflet molecules is preserved in SPMBs, suggesting that they are accessible on the surface of the bilayers. Such model membrane systems will be useful for many applications requiring detailed characterization of plasma membrane dynamics.

Membranes ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 30
Author(s):  
Erdinc Sezgin ◽  
Dario Carugo ◽  
Ilya Levental ◽  
Eleanor Stride ◽  
Christian Eggeling

Model membrane systems are essential tools for the study of biological processes in a simplified setting to reveal the underlying physicochemical principles. As cell-derived membrane systems, giant plasma membrane vesicles (GPMVs) constitute an intermediate model between live cells and fully artificial structures. Certain applications, however, require planar membrane surfaces. Here, we report a new approach for creating supported plasma membrane bilayers (SPMBs) by bursting cell-derived GPMVs using ultrasound within a microfluidic device. We show that the mobility of outer leaflet molecules is preserved in SPMBs, suggesting that they are accessible on the surface of the bilayers. Such model membrane systems are potentially useful in many applications requiring detailed characterization of plasma membrane dynamics.


2017 ◽  
Vol 28 (11) ◽  
pp. 1507-1518 ◽  
Author(s):  
Falk Schneider ◽  
Dominic Waithe ◽  
Mathias P. Clausen ◽  
Silvia Galiani ◽  
Thomas Koller ◽  
...  

Diffusion and interaction dynamics of molecules at the plasma membrane play an important role in cellular signaling and are suggested to be strongly associated with the actin cytoskeleton. Here we use superresolution STED microscopy combined with fluorescence correlation spectroscopy (STED-FCS) to access and compare the diffusion characteristics of fluorescent lipid analogues and GPI-anchored proteins (GPI-APs) in the live-cell plasma membrane and in actin cytoskeleton–free, cell-derived giant plasma membrane vesicles (GPMVs). Hindered diffusion of phospholipids and sphingolipids is abolished in the GPMVs, whereas transient nanodomain incorporation of ganglioside lipid GM1 is apparent in both the live-cell membrane and GPMVs. For GPI-APs, we detect two molecular pools in living cells; one pool shows high mobility with transient incorporation into nanodomains, and the other pool forms immobile clusters, both of which disappear in GPMVs. Our data underline the crucial role of the actin cortex in maintaining hindered diffusion modes of many but not all of the membrane molecules and highlight a powerful experimental approach to decipher specific influences on molecular plasma membrane dynamics.


2012 ◽  
Vol 33 (7) ◽  
pp. 535-542 ◽  
Author(s):  
Wenjun J. Sun ◽  
Mehri Kaviani Mogadam ◽  
Marianne Sommarin ◽  
Henrietta Nittby ◽  
Leif G. Salford ◽  
...  

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