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1.
Infrared spectroscopy is a powerful technique for examining lipid bilayers; however, it says little about the fluidity of the bilayer-a key physical aspect. It is shown here that it is possible to both acquire spectroscopic data of supported lipid bilayer samples and make measurements of the membrane fluidity. Attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FT-IR) is used to obtain the spectroscopic information and fluorescence recovery after photobleaching (FRAP) is used to determine the fluidity of the samples. In the infrared spectra of lipid bilayers composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine, the following major peaks were observed; nu(as)(CH3) 2954 cm(-1), nu(s)(CH3) 2870 cm(-1), nu(as)(CH2) 2924 cm(-1), nu(s)(CH2) 2852 cm(-1), nu(C=O) 1734 cm(-1), delta(CH2) 1463-1473 cm(-1), nu(as)(PO2-) 1226 cm(-1), nu(s)(PO2-) 1084 cm(-1), and nu(as)(N+(CH3)3) 973 cm(-1). The diffusion coefficient of the same lipid bilayer was measured to be 3.5 +/- 0.5 micom(2)/s with visual recovery also noted through use of epifluorescence microscopy. FRAP and visual data confirm the formation of a uniform, mobile supported lipid bilayer. The combination of ATR-FT-IR and FRAP provides complementary data giving a more complete picture of fully hydrated model membrane systems.  相似文献   

2.
Supported lipid bilayers (SLBs) at surfaces provide a route to quantitatively study molecular interactions with and at lipid membranes via different surface-based analytical techniques. Here, a method to fabricate SLBs with controlled curvatures, in the nanometer regime over large areas, is presented, utilizing lipid vesicle rupture onto nanostructured sensor substrates. Heat treated colloidal particle masks were used as templates to produce silicon dioxide films with systematically varied radius of curvature (ROC, 70 to 170 nm are demonstrated) and quartz crystal microbalance with dissipation monitoring (QCM-D) was used to confirm vesicle rupture onto such structured surfaces. Fluorescence microscopy was used to show fluidity of the supported membranes. The formation of confluent SLBs is demonstrated at the nanostructured surfaces from vesicles composed of POPC lipids. However, at surfaces with decreasing ROCs, vesicle rupture was hindered but with an increasing fraction of the positively charged lipid POEPC in the vesicles, it was possible to form good quality supported bilayers on all curvatures studied. Curved SLBs open up the possibility to systematically study the influence of curvature on molecular interactions at lipid membranes.  相似文献   

3.
Liu KW  Biswal SL 《Analytical chemistry》2011,83(12):4794-4801
The interaction of surfactants with lipid membranes can result in composition change, area expansion, solubilization, or the formation of protrusion features of the membranes. Amphipathic surfactant molecules are simplified analogues to membrane-active drugs and peptides which are known for inserting into lipid bilayers; however, the effect of these amphipathic molecules on supported membranes is not well characterized. In this paper we explore the use of microcantilever sensors to quantify surfactants' effects on lipid membranes. We use microcantilevers which are coated with lipid membranes to probe the interactions between lysolipids and supported lipid bilayers (SLBs). In particular, we investigate the effects of four zwitterionic surfactants similar to phospholipids: lysolipids of different aliphatic chain lengths (lysophosphocholines, lysoPCs, 12:0, 14:0, 16:0, and 18:0) on 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine-supported lipid bilayers. By monitoring the deflection of the microcantilevers, real-time free energy changes in the SLBs upon the addition of lysolipids can be detected. Additionally, the bending direction reveals whether the lysoPCs incorporate into or solubilize the SLB. When the bulk lysoPC concentration is less than its critical micelle concentration (CMC), we observe a compressive bending of the microcantilever, indicating adsorption to the SLB. Additionally, the change in surface stress is found to be proportional to the amount of membrane-bound lysoPCs. For bulk concentrations greater than the CMC, lysoPCs 12:0 and 14:0, there is tensile bending, indicating that the lysoPCs begin to solubilize and destroy the SLBs. Interestingly, this is not observed for lysoPCs with longer chain lengths. This new method of using microcantilevers for detecting and quantifying the surfactant insertion and solubilization of SLBs offers additional insights into the interactions between small amphipathic molecules and lipid membranes.  相似文献   

4.
Model biomembranes composed of poly-DATT/DGS/POPA and poly-DATT/DGS/CL were separately prepared on gold electrodes. A monolayer of 1,2-dioleoyl-sn-glycero-3-succinate (DGS) was covalently bonded onto electrochemically grown poly-(3,4-diamiono-2,2:5,2-terthiophene) (DATT) layers (thickness of approximately 300 nm; particle size of approximately 50 to 70 nm). The numbers of unit molecules of the poly-DATT layer and of the DGS immobilized onto the poly-DATT layers were 1.53 x 10(-7) and 1.56 x 10(-9) mol cm(-2), respectively, using a quartz crystal microbalance technique. The lipid bilayers of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphate (POPA) and cardiolipin (CL) were formed onto the poly-DATT/DGS layer using the Langmuir-Blodgett technique. The surface characterizations of each step were investigated by SEM, AFM, and XPS analyses. Cytochrome c (cyt c) was immobilized onto these model biomembranes through the charge interaction between the positive charges of cyt c and the negative charges of phosphate groups in CL or POPA lipids. At the POPA- and CL-modified biomembranes, the formal potentials of the redox couple of the immobilized cyt c were 0.22 and 0.23 V (vs Ag/AgCl), respectively. The redox reaction of the immobilized cyt c at the POPA- and CL-modified biomembranes was quasireversible, and the electron-transfer rate constants were 0.121 s(-1) and 0.133 s(-1), respectively. The applicability of these cyt c immobilized bioimitation membranes as the biosensors was tested for the determination of superoxide.  相似文献   

5.
Curved lipid membranes are ubiquitous in living systems and play an important role in many biological processes. To understand how curvature and lipid composition affect membrane formation and fluidity, we have assembled and studied mixed 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) supported lipid bilayers on amorphous silicon nanowires grown around carbon nanotube cores with controlled wire diameters ranging from 20 to 200 nm. We found that lipid vesicles fused onto nanowire substrates and formed continuous bilayers for all DOPC-DOPE mixtures tested (with the DOPE content of up to 30%). Our measurements demonstrate that nanowire-supported bilayers are mobile, exhibit fast recovery after photobleaching, and have a low concentration of defects. Lipid diffusion coefficients in these high-curvature tubular membranes are comparable to the values reported for flat supported bilayers and increase slightly with decreasing nanowire diameter. A free space diffusion model adequately describes the effect of bilayer curvature on the lipid mobility for nanowire substrates with diameters greater than 50 nm, but shows significant deviations from the experimental values for smaller diameter nanowires.  相似文献   

6.
Chiral second harmonic generation (C-SHG) has been used for the label-free detection of (R)-(+)-1,1'-bi-2-naphthol (RBN) and (S)-(+)-1,1'-bi-2-naphthol (SBN) binding to planar-supported lipid bilayers of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphotidylcholine (POPC) based on the intrinsic chirality of the molecules. C-SHG adsorption isotherms of RBN and SBN reveal Langmuir adsorption behavior with binding constants of 2.7 +/- 0.2 x 10(5) M(-1) and 3.0 +/- 0.1 x 10(5) M(-1), respectively. The kinetics of RBN binding to a POPC bilayer was also measured. It was determined that the adsorption rate for RBN was 5.7 +/- 0.4 x 10(3) s(-1)M(-1) and the desorption rate was 2.1 +/- 0.8 x 10(-2) s(-1). From the kinetic data a binding constant of 2.7 +/- 1.0 x 10(5) M(-1) was calculated, which agrees well with the thermodynamic measurement. The C-SHG technique was correlated with surface tension measurements in order to determine the RBN surface excess within the POPC membrane. The maximum surface excess of RBN in a monolayer of POPC was 4.3 +/- 0.5 x 10(-11) mol cm2. Using the maximum surface excess in conjunction with the C-SHG binding data a lower limit of detection of 1.5 +/- 0.1 x 10(-13) mols cm(-2) was calculated. The results of these studies show that C-SHG is a powerful tool for the study of chiral molecular interactions at surfaces.  相似文献   

7.
We have successfully synthesized a lipid containing the pyranine dye as the hydrophilic headgroup. This lipid was incorporated into liposomes with 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine as the major component. The resultant liposomes displayed differential modulations in fluorescence emission intensity in the presence of nanomolar concentrations of different glycosaminoglycans. Linear discriminant analysis of the fluorescence response data demonstrate that the liposomes are able to distinguish between different GAGs. In addition, we also demonstrate that the liposomes incorporating the pyranine lipid are able to distinguish between dilute serum from healthy individuals and serum containing elevated chondroitin sulfate (simulated serum from an Alzheimer's disease patient).  相似文献   

8.
Here we report the use of counter-propagating second harmonic generation (SHG) to image the interactions between the local anesthetic tetracaine and a multicomponent planar supported lipid bilayer array in a label-free manner. The lipid bilayer arrays, prepared using a 3D continuous flow microspotter, allow the effects of lipid phase and cholesterol content on tetracaine binding to be examined simultaneously. SHG images show that tetracaine has a higher binding affinity to liquid-crystalline phase lipids than to solid-gel phase lipids. The presence of 28 mol % cholesterol decreased the binding affinity of tetracaine to bilayers composed of the mixed chain lipid, 1-steroyl-2-oleoyl-sn-glycero-3-phophocholine (SOPC), and the saturated lipids 1,2-dimyristoyl-sn-glycero-3-phophocholine (DMPC) and 1,2-dipamitoyl-sn-glycero-3-phophocholine (DPPC) while having no effect on diunsaturated 1,2-dioleoyl-sn-glycero-3-phophocholine (DOPC). The maximum surface excess of tetracaine increases with the degree of unsaturation of the phospholipids and decreases with cholesterol in the lipid bilayers. The paper demonstrates that SHG imaging is a sensitive technique that can directly image and quantitatively measure the association of a drug to a multicomponent lipid bilayer array, providing a high-throughput means to assess drug-membrane interactions.  相似文献   

9.
Keffer CE  Torr MR  Zukic M  Spann JF  Torr DG  Kim J 《Applied optics》1994,33(25):6041-6045
Advances in vacuum ultraviolet thin-film filter technology have been made through the use of filter designs with multilayers of materials such as Al(2)O(3), BaF(2), CaF(2), HfO(2), LaF(3), MgF(2), and SiO(2). Our immediate application for these filters will be in an imaging system to be flown on a satellite where a 2 × 9 R(E) orbit will expose the instrument to approximately 250 krad of radiation. Because to our knowledge no previous studies have been made on the potential radiation damage of these materials in the thin-film format, we report on such an assessment here. Transmittances and reflectances of BaF(2), CaF(2), HfO(2), MgF(2), and SiO(2) thin films on MgF(2) substrates, Al(2)O(3) thin films on fused-silica substrates, uncoated fused silica and MgF(2), and four multilayer filters made from these materials were measured from 120 to 180 nm beforeand after irradiation by 250 krad from a (60)Co gamma radiation source. No radiation-induced losses in transmittance or reflectance occurred in this wavelength range. Additional postradiation measurements from 160 to 300 nm indicates 2-5% radiation-induced absorption near 260 nm in some of the samples with MgF(2) substrates. From these measurements we conclude that far-ultraviolet filters made from the materials tested should experience less than 5% change from exposure to up to 250 krad of high-energy radiation in space applications.  相似文献   

10.
The controlled fabrication of biocompatible devices made of lipid bilayers deposited onto flat solid supports presents interest as models of cell membranes as well as for their biotechnological applications. We report here on the formation of supported lipid bilayers on silica nanoparticles (nanoSLBs). The successive steps of the adsorption of lipid vesicles on nanoparticles and the formation of nanoSLBs are revealed in detail by cryotransmission electron microscopy (cryo-EM). The formation of nanoSLBs was achieved for liposomes with positive, neutral, and low net negative charge, while liposomes with a high net negative charge adsorbed to silica nanoparticles but did not rupture. The nanoSLBs were found to follow faithfully the surface contours of the particles, information yet unavailable for SLB formation on planar solid substrates.  相似文献   

11.
Bio-interfaces such as bio-membranes are of outmost importance for a variety of live processes. Among them are cell-interactions which take place in, on or through cell membranes. Therefore we propose to cover metallic surfaces with phospholipids to facilitate cell-material interaction. Four lipids, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), 1-palmitoyl-2- oleoyl-sn-glycero-3-[phospho-L-serine] (POPS) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-rac-(1-glycerol) (POPG), were applied to four metallic growth substrates with different surface structure, roughness and porosity. The interaction of the osteosarcoma cell line MG-63 was investigated in terms of cell adhesion and viability (MTT (methylthiazolyldiphenyl-tetrazolium bromide) assay). While POPS in general had a negative influence, the most suitable combination in terms of viability per adherent MG-63 is the coating of porous Ti6Al4V material with the phospholipids POPE or POPC. The analysis of viability of mouse macrophages RAW 264.7 and their tumor necrosis factor α (TNF-α) release showed that the adhesion and viability is worst on POPS while the TNF-α release was highest. To elucidate the potential of phospholipids to prevent or support bacterial growth, the bacterial number of Gram positive and Gram negative bacteria was investigated. For lipid concentrations higher than 1 mM in solution a growth stimulating effect independent of the lipid type was detected. On a lipid coated surface the number of bacteria was reduced by 81%, 74% and 51% for POPC, POPG and POPE.  相似文献   

12.
We report a simple solution process to form controlled patterns of aligned single-walled carbon nanotubes on solid substrates. The essential step of the process is to deposit a dilute solution of DNA-wrapped carbon nanotubes (DNA-CNTs) on a SiO(2) surface covered with a thin hydrophobic layer. This leads to deposition of fully aligned CNTs. The alignment pattern can be controlled by metal electrodes in the deposition region and can be quantitatively modeled by the behavior of a quasi-two-dimensional DNA-CNT nematic phase near the solution/SiO(2) interface. These results point to the possibility of rational design and economical fabrication of CNT alignment patterns on solid substrates.  相似文献   

13.
The influence of layer-by-layer films of polyaniline and Ni-tetrasulfonated phthalocyanine (PANI/Ni-TS-Pc) on the electrical performance of polymeric light-emitting diodes (PLED) made from (poly[2-methoxy-5-(2′-ethyl-hexyloxy)-1,4-phenylene vinylene]) (MEH-PPV) is investigated by using current versus voltage measurements and impedance spectroscopy. The PLED is composed by a thin layer of MEH-PPV sandwiched between indium tin oxide (ITO) and aluminum electrodes, resulting in the device structure ITO/(PANI/Ni-TS-Pc)n/MEH-PPV/Al, where n stands for the number of PANI/Ni-TS-Pc bilayers. The deposition of PANI/Ni-TS-Pc leads to a decrease in the driving voltage of the PLEDs, which reaches a minimum when n = 5 bilayers. In addition, impedance spectroscopy data reveal that the PLED impedance decreases as more PANI/Ni-TS-Pc bilayers are deposited. The PLED structure is further described by an equivalent circuit composed by two R-C combinations, one for the bulk and other for the interface components, in series with a resistance originated in the ITO contact. From the impedance curves, the values for each circuit element is determined and it is found that both, bulk and interface resistances are decreased upon PANI/Ni-TS-Pc deposition. The results indicate that PANI/Ni-TS-Pc films reduce the contact resistance at ITO/MEH-PPV interface, and for that reason improve the hole-injection within the PLED structure.  相似文献   

14.
Nanoparticles with widely varying physical properties and origins (spherical versus irregular, synthetic versus biological, organic versus inorganic, flexible versus rigid, small versus large) have been previously noted to translocate across the cell plasma membrane. We have employed atomic force microscopy to determine if the physical disruption of lipid membranes, formation of holes and/or thinned regions, is a common mechanism of interaction between these nanoparticles and lipids. It was found that a wide variety of nanoparticles, including a cell penetrating peptide (MSI-78), a protein (TAT), polycationic polymers (PAMAM dendrimers, pentanol-core PAMAM dendrons, polyethyleneimine, and diethylaminoethyl-dextran), and two inorganic particles (Au-NH2, SiO2-NH2), can induce disruption, including the formation of holes, membrane thinning, and/or membrane erosion, in supported lipid bilayers.  相似文献   

15.
A prototype amperometric fructose biosensor based on membrane-bound fructose dehydrogenase (Gluconobacter sp.) and the coenzyme ubiquinone-6 immobilized in a membrane mimetic layer on a gold electrode has been constructed and tested. A bare gold electrode first was modified through chemisorption of a mixture of octadecyl mercaptan and two short-chain disulfides, 3,3'-dithiodipropionic acid and cystamine dihydrochloride. The membrane-bound enzyme, coenzyme, and additional phospholipid were codeposited through a detergent dialysis protocol. The short-chain modifiers may provide electrostatic interactions with enzyme surface charges, while the alkanethiolate and phospholipids enable hydrophobic interaction with the largely lipophilic, membrane-bound enzyme. At oxidizing potentials, the enzyme electrode responded with catalytic current densities up to 45 μA/cm(2) when exposed to fructose at 10 mM. The sensor exhibited a response time of less than 20 s, a sensitivity of 15 μA/cm(2)·mM and a detection limit of less than 10 μM. Biosensor measurements of d-fructose in apple and orange juice agreed to within a few percent with those made with an enzymatic spectrophotometric assay. The membrane mimetic layer effectively blocked access of interfering ascorbic acid to the electrode surface. Only a 4% positive error was observed in the presence of ascorbic acid at 5% of the fructose concentration (2 mM), which indicates that this construct could be particularly useful for quantitation of fructose in citrus juice.  相似文献   

16.
SiO2在ZnO—Bi2O3—Sb2O3—BaO系压敏陶瓷中的作用   总被引:1,自引:0,他引:1  
SiO2添加剂对ZnO-Bi2O3-Sb2O3-BaO系和通常的ZnO-Bi2O3-Sb2O3系压敏陶瓷电性能的作用和影响有很大不同。通过小电流区伏安特性,交变电压下复阻抗特性,以及对复电容平面图中半弧特性的分析可知,SiO2对ZnO-Bi2O3-Sb2O3-BaO系压敏陶瓷中电性能影响是晶界组分及微结构变化造成的。  相似文献   

17.
Multilayered Ge nanocrystals embedded in Si and Ge oxide films have been fabricated on Si?substrate by a (SiO(2)+Ge)/(SiO(2)+GeO(2)) superlattice approach, using an rf magnetron sputtering technique with a Ge+SiO(2) composite target and subsequent thermal annealing in N(2) ambient at 750?°C for 5?min. X-ray diffraction (XRD) measurements indicated the formation of Ge nanocrystals with an average size estimated to be 9.8?nm. Raman scattering spectra showed a peak of the Ge-Ge vibrational mode shifted downwards to 298.8?cm(-1), which was caused by quantum confinement of phonons in the Ge nanocrystals. X-ray photoemission spectroscopy (XPS) analysis demonstrated that the Ge chemical state is mainly Ge(0) in the (SiO(2)+Ge) layer and Ge(4+) in the (SiO(2)+GeO(2)) layer in the superlattice structure. Transmission electron microscopy (TEM) revealed that Ge nanocrystals were confined in (SiO(2)+Ge) layers, and had good crystallinity. This superlattice approach significantly improved both the size uniformity of Ge nanocrystals and their uniformity of spacing on the 'Z' growth direction compared with the conventional Ge-ncs fabrication method using a single and thick SiO(2) matrix film.  相似文献   

18.
The effect of a polyelectrolyte (PE) multilayers made by a layer-by-layer technique on the response of a quartz crystal microbalance (QCM) is studied by using novel mathematical methods based on the M?bius transformations and their matrix representations in the complex plane. In the first method, the basic properties of the M?bius transformation are used for obtaining the PE bilayer matrix from the QCM impedance measurements taken at four different numbers of layers. In the second method, nonlinear fitting with concomitant error estimation is used for obtaining the elements of the bilayer matrix. The methods are applied to a multilayer composed of 150 bilayers of poly(sodium 4-styrenesulfonate) and poly(diallyldimethylammonium) chloride on a quartz crystal resonator. The structure of the system is discussed, and the bulk acoustic impedance and areal mass density of the bilayer are calculated from the layer matrix.  相似文献   

19.
Goswami  Ankur  Dhandaria  Priyesh  Pal  Soupitak  McGee  Ryan  Khan  Faheem  Antić  Željka  Gaikwad  Ravi  Prashanthi  Kovur  Thundat  Thomas 《Nano Research》2017,10(10):3571-3584
This study reports on the mid-infrared (mid-IR) photothermal response of multilayer MoS2 thin films grown on crystalline (p-type silicon and c-axisoriented single crystal sapphire) and amorphous (Si/SiO2 and Si/SiN) substrates by pulsed laser deposition (PLD).The photothermal response of the MoS2 films is measured as the changes in the resistance of the MoS2 films when irradiated with a mid-IR (7 to 8.2 μm) source.We show that enhancing the temperature coefficient of resistance (TCR) of the MoS2 thin films is possible by controlling the film-substrate interface through a proper choice of substrate and growth conditions.The thin films grown by PLD are characterized using X-ray diffraction,Raman,atomic force microscopy,X-ray photoelectron microscopy,and transmission electron microscopy.The high-resolution transmission electron microscopy (HRTEM) images show that the MoS2 films grow on sapphire substrates in a layer-by-layer manner with misfit dislocations.The layer growth morphology is disrupted when the films are grown on substrates with a diamond cubic structure (e.g.,silicon) because of twin growth formation.The growth morphology on amorphous substrates,such as Si/SiO2 or Si/SiN,is very different.The PLD-grown MoS2 films on silicon show higher TCR (-2.9% K-1 at 296 K),higher mid-IR sensitivity (△R/R =5.2%),and higher responsivity (8.7 V·W-1) compared to both the PLD-grown films on other substrates and the mechanically exfoliated MoS2 flakes transferred to different substrates.  相似文献   

20.
The development of Ni/Au contacts to Mg-doped GaN nanowires (NWs) is examined. Unlike Ni/Au contacts to planar GaN, current-voltage (I-V) measurements of Mg-doped nanowire devices frequently exhibit a strong degradation after annealing in N(2)/O(2). This degradation originates from the poor wetting behavior of Ni and Au on SiO(2) and the excessive void formation that occurs at the metal/NW and metal/oxide interfaces. The void formation can cause cracking and delamination of the metal film as well as reduce the contact area at the metal/NW interface, which increases the resistance. The morphology and composition of the annealed Ni/Au contacts on SiO(2) and the p-GaN films were investigated by scanning electron microscopy (SEM), energy-dispersive x-ray spectroscopy (EDS) and x-ray diffraction (XRD) measurements. Adhesion experiments were performed in order to determine the degree of adhesion of the Ni/Au films to the SiO(2) as well as observe and analyze the morphology of the film's underside by SEM. Device degradation from annealing was prevented through the use of a specific adhesion layer of Ti/Al/Ni deposited prior to the nanowire dispersal and Ni/Au deposition. I-V measurements of NW devices fabricated using this adhesion layer showed a decrease in resistance after annealing, whereas all others showed an increase in resistance. Transmission electron microscopy (TEM) on a cross-section of a NW with Ni/Au contacts and a Ti/Al/Ni adhesion layer showed a lack of void formation at the contact/NW interface. Results of the XRD and TEM analysis of the NW contact structure using a Ti/Al/Ni adhesion layer suggests Al alloying of the Ni/Au contact increases the adhesion and stability of the metal film as well as prevents excessive void formation at the contact/NW interface.  相似文献   

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