Measurement of membrane tension of free standing lipid bilayers via laser-induced surface deformation spectroscopy

Soft Matter ◽  
2015 ◽  
Vol 11 (44) ◽  
pp. 8641-8647 ◽  
Author(s):  
Tomohiko Takei ◽  
Tatsuya Yaguchi ◽  
Takuya Fujii ◽  
Tomonori Nomoto ◽  
Taro Toyota ◽  
...  

Non-invasive measurement of the membrane tension of free-standing black lipid membranes (BLMs), with sensitivity on the order of μN m−1, was achieved using laser-induced surface deformation (LISD) spectroscopy.

2018 ◽  
Vol 34 (11) ◽  
pp. 1237-1242 ◽  
Author(s):  
Tomonori NOMOTO ◽  
Masahiro TAKAHASHI ◽  
Takuya FUJII ◽  
Luca CHIARI ◽  
Taro TOYOTA ◽  
...  

Author(s):  
Neng-Bo He ◽  
S.W. Hui

Monolayers and planar "black" lipid membranes have been widely used as models for studying the structure and properties of biological membranes. Because of the lack of a suitable method to prepare these membranes for electron microscopic observation, their ultrastructure is so far not well understood. A method of forming molecular bilayers over the holes of fine mesh grids was developed by Hui et al. to study hydrated and unsupported lipid bilayers by electron diffraction, and to image phase separated domains by diffraction contrast. We now adapted the method of Pattus et al. of spreading biological membranes vesicles on the air-water interfaces to reconstitute biological membranes into unsupported planar films for electron microscopic study. hemoglobin-free human erythrocyte membrane stroma was prepared by hemolysis. The membranes were spreaded at 20°C on balanced salt solution in a Langmuir trough until a surface pressure of 20 dyne/cm was reached. The surface film was repeatedly washed by passing to adjacent troughs over shallow partitions (fig. 1).


Antibiotics ◽  
2019 ◽  
Vol 8 (1) ◽  
pp. 12 ◽  
Author(s):  
Amani Alghalayini ◽  
Alvaro Garcia ◽  
Thomas Berry ◽  
Charles Cranfield

This review identifies the ways in which tethered bilayer lipid membranes (tBLMs) can be used for the identification of the actions of antimicrobials against lipid bilayers. Much of the new research in this area has originated, or included researchers from, the southern hemisphere, Australia and New Zealand in particular. More and more, tBLMs are replacing liposome release assays, black lipid membranes and patch-clamp electrophysiological techniques because they use fewer reagents, are able to obtain results far more quickly and can provide a uniformity of responses with fewer artefacts. In this work, we describe how tBLM technology can and has been used to identify the actions of numerous antimicrobial agents.


2021 ◽  
Vol 118 (48) ◽  
pp. e2113202118
Author(s):  
Rafael L. Schoch ◽  
Frank L. H. Brown ◽  
Gilad Haran

Lipid membranes are complex quasi–two-dimensional fluids, whose importance in biology and unique physical/materials properties have made them a major target for biophysical research. Recent single-molecule tracking experiments in membranes have caused some controversy, calling the venerable Saffman–Delbrück model into question and suggesting that, perhaps, current understanding of membrane hydrodynamics is imperfect. However, single-molecule tracking is not well suited to resolving the details of hydrodynamic flows; observations involving correlations between multiple molecules are superior for this purpose. Here dual-color molecular tracking with submillisecond time resolution and submicron spatial resolution is employed to reveal correlations in the Brownian motion of pairs of fluorescently labeled lipids in membranes. These correlations extend hundreds of nanometers in freely floating bilayers (black lipid membranes) but are severely suppressed in supported lipid bilayers. The measurements are consistent with hydrodynamic predictions based on an extended Saffman–Delbrück theory that explicitly accounts for the two-leaflet bilayer structure of lipid membranes.


1995 ◽  
Vol 50 (11-12) ◽  
pp. 833-839 ◽  
Author(s):  
Alexander Pazur

Abstract Black lipid membranes were prepared on a Teflon septum separating electrically the two chambers of a Teflon cuvette, using the technique of Mueller et al., (Nature 194. 979 (1962)). An external, static magnetic field was applied, whose intensity varied from 0 G to 100 G at the membrane location. Field applications higher than 10 G are effecting higher leakage currents, increased capacity and faster breakdown of the bilayer state, as compared to the absence of a magnetic field. If bilayers were doped with chlorophyll a, these effects were increased. Quantum mechanical and thermodynamical phenomena on membranes will be discussed as possible origins of these effects.


2011 ◽  
Vol 137 (2) ◽  
pp. 217-223 ◽  
Author(s):  
Sourabh Banerjee ◽  
Crina M. Nimigean

Discoidal lipoproteins are a novel class of nanoparticles for studying membrane proteins (MPs) in a soluble, native lipid environment, using assays that have not been traditionally applied to transmembrane proteins. Here, we report the successful delivery of an ion channel from these particles, called nanoscale apolipoprotein-bound bilayers (NABBs), to a distinct, continuous lipid bilayer that will allow both ensemble assays, made possible by the soluble NABB platform, and single-molecule assays, to be performed from the same biochemical preparation. We optimized the incorporation and verified the homogeneity of NABBs containing a prototypical potassium channel, KcsA. We also evaluated the transfer of KcsA from the NABBs to lipid bilayers using single-channel electrophysiology and found that the functional properties of the channel remained intact. NABBs containing KcsA were stable, homogeneous, and able to spontaneously deliver the channel to black lipid membranes without measurably affecting the electrical properties of the bilayer. Our results are the first to demonstrate the transfer of a MP from NABBs to a different lipid bilayer without involving vesicle fusion.


2021 ◽  
Vol 118 (31) ◽  
pp. e2104610118
Author(s):  
Jean-Baptiste Fleury ◽  
Vladimir A. Baulin

Estimated millions of tons of plastic are dumped annually into oceans. Plastic has been produced only for 70 y, but the exponential rise of mass production leads to its widespread proliferation in all environments. As a consequence of their large abundance globally, microplastics are also found in many living organisms including humans. While the health impact of digested microplastics on living organisms is debatable, we reveal a physical mechanism of mechanical stretching of model cell lipid membranes induced by adsorbed micrometer-sized microplastic particles most commonly found in oceans. Combining experimental and theoretical approaches, we demonstrate that microplastic particles adsorbed on lipid membranes considerably increase membrane tension even at low particle concentrations. Each particle adsorbed at the membrane consumes surface area that is proportional to the contact area between particle and the membrane. Although lipid membranes are liquid and able to accommodate mechanical stress, the relaxation time is much slower than the rate of adsorption; thus, the cumulative effect from arriving microplastic particles to the membrane leads to the global reduction of the membrane area and increase of membrane tension. This, in turn, leads to a strong reduction of membrane lifetime. The effect of mechanical stretching of microplastics on living cells membranes was demonstrated by using the aspiration micropipette technique on red blood cells. The described mechanical stretching mechanism on lipid bilayers may provide better understanding of the impact of microplastic particles in living systems.


2019 ◽  
Vol 19 (11) ◽  
pp. 7149-7155
Author(s):  
Shengwei Tan ◽  
Ling Zhang ◽  
Lijuan Yu ◽  
Lei Xu

Integrated nanopores are novel and versatile single-molecule sensors for individual label-free biopolymer detection and characterization. However, their studies and application requires a stable lipid bilayer to maintain protein function. Herein, we describe a method for producing lipid bilayers across a nanopore array on a silicon nitride substrate. We used a painting technique commonly used with Teflon films to embed α-hemolysin (α-HL) into bilayer lipid membranes (BLMs) to form an ion channel. This was carried out in nanofluid developed in our lab. The membrane formation process, stability of BLMs and ion channel recordings were monitored by patch clamp in real-time. BLM formation was demonstrated by electrical recording (<10 pS conductance) of suspended lipid bilayers spanning a nanopore in the range of ±100 mV. Membrane resistance (Rm) and capacitance (Cm) of the device with the bilayer were assessed by membrane test as above 1.0 GΩ and ~20±2 pF, respectively. The silicon nitride surface and aperture edge were smooth at the nanometer lever leading to remarkable membrane stability. The membrane lifetime was 5–24 h. A single α-HL channel inserted in 30–60 min applied a potential of +100 mV. The α-HL channel currents were recorded at ~100±10 pA. Such integrated nanopores enable analysis of channel functions under various solution conditions from the same BLM. This will open up a variety of applications for ion channels including high-throughput medical screening and diagnosis.


2016 ◽  
Author(s):  
◽  
Matthew McCune

This dissertation presents two research projects that apply theoretical and computational modeling to (1) describe and predict the formation and shape evolution of three-dimensional (3D) bioprinted tissue constructs, and (2) investigate the effect of a silica substrate on the structural and dynamic properties of a single fully hydrated lipid bilayer. (1) Bioprinting, a novel tissue engineering technique, has the ultimate goal of using 3D printers with bioink made from a person’s own cells to create tissues in the laboratory for transplantation or drug testing. The outcome of the post-bioprinting process, where the bioink particles fuse to form the desired 3D tissue construct, is difficult to predict and experimental techniques have generally been optimized through trial and error. To address this shortcoming, by employing theoretical modeling and computer simulations, we have developed and implemented an effective procedure that is capable of describing and predicting the shape dynamics during post-printing structure formation in 3D bioprinting. In particular, we have explained and demonstrated that the post-printing fusion process is considerably faster when using cylindrical instead of spheroidal bioink particles, a result that has considerable practical implication for extrusion bioprinting. (2) The study of lipid bilayers using neutron scattering experiments requires samples that contain a large stack of membranes. The analysis and computer simulation of such systems is challenging mainly due to the unknown amount of water separating the membranes. To overcome this difficulty, more recent experiments place single lipid membranes onto a support and stack about a hundred of them together. In this project we use molecular dynamics simulations of both free-standing and hydrated single-supported lipid bilayers to investigate the effect of the silica substrate on the structural and dynamical properties of the lipids and hydration waters. Our results may provide useful information in interpreting some recent neutron scattering experiments.


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