Refined contour analysis of giant unilamellar vesicles

The fluctuation spectrum of giant unilamellar vesicles is measured using a high-resolution contour detection technique. An analysis at higher q vectors than previously achievable is now possible due to technical improvements of the experimental setup and of the detection algorithm. The global fluctuation spectrum is directly fitted to deduce the membrane tension and the bending modulus of lipid membranes. Moreover, we show that the planar analysis of fluctuations is valid for spherical objects, even at low wave vectors. Corrections due to the integration time of the video camera and to the section of a 3D object by the observation plane are introduced. A precise calculation of the error bars has been done in order to provide reliable error estimate. Eventually, using this technique, we have measured bending moduli for EPC, SOPC and SOPC: CHOL membranes confirming previously published values. An interesting application of this technique can be the measurement of the fluctuation spectra for non-equilibrium membranes, such as active membranes.Received: 11 March 2004, Published online: 21 April 2004PACS: 87.16.Dg Membranes, bilayers and vesicles - 05.40.-a Fluctuation phenomena, random processes, noise and Brownian motion - 07.60.Pb Conventional optical microscopes     

Transition from small to big charged unilamellar vesicles

The transition from small to big unilamellar vesicles predicted by a Poisson-Boltzmann Cell Model for the thermodynamics of a dilute phase of unilamellar charged vesicles is characterized. The radius as a function of experimental parameters is calculated and the coexistence region of small and big vesicles is identified. We further investigate the physical meaning in terms of simplified models, which allow for an identification of the role of parameters like the surface charge density or the Debye-length. Connections to experiments are discussed. Received: 20 October 1997 / Received in final form: 9 February 1998 / Accepted: 9 March 1998     

Regulation of the intermittent release of giant unilamellar vesicles under osmotic pressure

Osmotic pressure can break the fluid balance between intracellular and extracellular solutions. In hypo-osmotic solution, water molecules, which transfer into the cell and burst, are driven by the concentration difference of solute across the semi-permeable membrane. The complicated dynamic processes of intermittent bursts have been previously observed. However, the underlying physical mechanism has yet to be thoroughly explored and analyzed. Here, the intermittent release of inclusion in giant unilamellar vesicles was investigated quantitatively, applying the combination of experimental and theoretical methods in the hypo-osmotic medium. Experimentally, we adopted a highly sensitive electron multiplying charge-coupled device to acquire intermittent dynamic images. Notably, the component of the vesicle phospholipids affected the stretch velocity, and the prepared solution of vesicles adjusted the release time. Theoretically, we chose equations and numerical simulations to quantify the dynamic process in phases and explored the influences of physical parameters such as bilayer permeability and solution viscosity on the process. It was concluded that the time taken to achieve the balance of giant unilamellar vesicles was highly dependent on the molecular structure of the lipid. The pore lifetime was strongly related to the internal solution environment of giant unilamellar vesicles. The vesicles prepared in viscous solution were able to visualize long-lived pores. Furthermore, the line tension was measured quantitatively by the release velocity of inclusion, which was of the same order of magnitude as the theoretical simulation. In all, the experimental values well matched the theoretical values. Our investigation clarified the physical regulatory mechanism of intermittent pore formation and inclusion release, which provides an important reference for the development of novel technologies such as gene therapy based on transmembrane transport as well as controlled drug delivery based on liposomes.     

Influence of cholesterol on the bilayer properties of monounsaturated phosphatidylcholine unilamellar vesicles

The influence of cholesterol on the structure of unilamellar-vesicle (ULV) phospholipid bilayers is studied using small-angle neutron scattering. ULVs made up of short-, mid- and long-chain monounsaturated phospholipids (diCn :1PC, n = 14 , 18, 22, respectively) are examined over a range (0-45mol %) of cholesterol concentrations. Cholesterol's effect on bilayer structure is characterized through changes to the lipid's transmembrane thickness, lateral area and headgroup hydration. For all three lipids, analysis of the experimental data shows that the addition of cholesterol results in a monotonic increase of these parameters. In the case of the short- and mid-chain lipids, this is an expected result, however, such a finding was unexpected for the long-chain lipid. This implies that cholesterol has a pronounced effect on the lipid's hydrocarbon chain organization.     

电制备巨囊泡的方法和机理探索

利用倒置显微镜研究电制备巨囊泡,并分析其形成和成长的动力学机理.由于磷脂头部的电特性,电场对水化的磷脂双层的静电力促使磷脂膜内双层之间分离.在动力学的作用下,双层两叶的不对称受力引发其弯曲、出芽、膨胀、封闭以及相互融合.结果表明,电场参数、干磷脂膜的均匀性、缓冲液以及温度等因素影响巨囊泡的粒径、形状和稳定性.     

Adhesion of giant unilamellar vesicles on double-end grafted DNA carpets

We have recently shown that the bio-mimetic adhesion of Giant Unilamellar Vesicles on carpets of lambda-phage DNAs, grafted by one end to the substrate, leads to DNA scraping and stapling. As the lipid adhesion patch is built, outward forces stretch the DNA, while adhesion patch formation staples the chains into frozen conformations, trapped between the GUV membrane and the substrate. Analysis of the scraped and stapled DNA conformations provides a wealth of information about the membrane/polymer interactions at play during the formation of a bio-adhesive contact zone. In this paper we report new phenomena revealed by scraping and stapling phenomena associated with the bio-mimetic adhesion of Giant Unilamellar Vesicles on carpets of lambda-phage DNAs that were grafted to the substrate by both ends. In particular, the peculiar shapes of stapled DNA observed in this case, suggest that the membrane exerces not only outward radial forces during patch formation, but is is also able to confine the DNA molecules in the orthoradial direction.     

电制备巨囊泡的方法和机理探索

利用倒置显微镜研究电制备巨囊泡, 并分析其形成和成长的动力学机理. 由于磷脂头部的电特性, 电场对水化的磷脂双层的静电力促使磷脂膜内双层之间分离. 在动力学的作用下, 双层两叶的不对称受力引发其弯曲、出芽、膨胀、封闭以及相互融合. 结果表明, 电场参数、干磷脂膜的均匀性、缓冲液以及温度等因素影响巨囊泡的粒径、形状和稳定性.     

蜂毒肽浓度和磷脂组分对巨囊泡泄露的影响

蜂毒肽作为一种广谱抗菌肽已经得到广泛认知,用蜂毒肽构建载药体系攻击癌细胞研究正在兴起.基于仿生物膜模型探索其破坏机理,可以避免潜在活性细胞过程的影响.在此,我们选用细胞尺寸的单层巨囊泡膜模型,可在光学显微镜下直接观察和操作,获得仿正常细胞膜和仿癌细胞膜在不同蜂毒肽浓度刺激下的响应.研究得出,低浓度蜂毒肽诱导囊泡泄露实验表明中性磷脂囊泡以孔模式为主泄露,负电性磷脂囊泡以爆裂模式为主泄露;高浓度蜂毒肽诱导囊泡泄露实验表明负电性磷脂相较于中性磷脂可延迟蜂毒肽作用效果;蜂毒肽色氨酸残基荧光光谱表明囊泡膜表面蜂毒肽吸附量以及泄露模式依赖于磷脂组分.此外,推断了蜂毒肽对不同组分磷脂膜的破坏作用模型.研究为蜂毒肽在肿瘤细胞的作用机制及其衍生物的优化设计提供参考.     

Transition from unilamellar to bilamellar vesicles induced by an amphiphilic biopolymer

We report some unusual structural transitions upon the addition of an amphiphilic biopolymer to unilamellar surfactant vesicles. The polymer is a hydrophobically modified chitosan and it embeds its hydrophobes in vesicle bilayers. We study vesicle-polymer mixtures using small-angle neutron scattering (SANS) and cryotransmission electron microscopy (cryo-TEM). When low amounts of the polymer are added to unilamellar vesicles of ca. 120 nm diameter, the vesicle size decreases by about 50%. Upon further addition of polymer, lamellar peaks are observed in the SANS spectra at high scattering vectors. We show that these spectra correspond to a co-existence of unilamellar and bilamellar vesicles. The transition to bilamellar vesicles as well as the changes in unilamellar vesicle size are further confirmed by cryo-TEM. A mechanism for the polymer-induced transitions in vesicle morphology is proposed.     

Effect of intra- and extra-liposomal distribution of sodium chloride on the stability of large unilamellar vesicles

Three groups of 1,2-dipalmitoyl-rac-glycero-3-phosphocholine (DPPC) large unilamellar vesicle (LUV) dispersions were studied: LUV (A) dispersions with only extraliposomal sodium chloride (NaCl), LUV (B) dispersions with intra- and extraliposomal NaCl, and LUV (C) dispersions with only intraliposomal NaCl. The NaCl concentrations ranged from 0 to 150 mM. An abrupt increase in leakage was observed around -10 degree C for all the three groups of LUV, which coincided with the temperature of extraliposomal ice formation. Within the three groups, leakage of LUV (C) was significantly higher than the other groups. Extraliposomal ice formation and the resulting freeze-concentration of LUV may be the major cause of the leakage. Intraliposomal ice formation observed at -43 degree C seemed to stop leakage of LUV when LUV were frozen below -43 degree C. An exotherm of eutectic crystallization of NaCl was occasionally observed at -37 degree C, with a higher probability of formation at 150 mM extraliposomal NaCl than at 50 mM. The eutectic crystals were thought to cause additional leakage from the LUV (B).     

Tilt angle of lipid acyl chains in unilamellar vesicles determined by ellipsometric light scattering

Ellipsometric light scattering (ELS) at room temperature is applied to unilamellar vesicles (~50 nm radius) of 1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) in the gel phase and of 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC) in the liquid-crystaline phase. A high sensitivity of this technique to the local anisotropy is found. From the resulting local birefringence, a lower limit of (29 ±0.5)^○ for the average tilt angle of the lipid chains of DPPC with respect to the membrane normal is estimated. This tilt angle value is slightly lower than literature values for the tilt angle in oriented lipid multi-bilayers on solid substrates.     

Dynamics of viscous vesicles in shear flow

The dynamics of giant lipid vesicles under shear flow is experimentally investigated. Consistent with previous theoretical and numerical studies, two flow regimes are identified depending on the viscosity ratio between the interior and the exterior of the vesicle, and its reduced volume or excess surface. At low viscosity ratios, a tank-treading motion of the membrane takes place, the vesicle assuming a constant orientation with respect to the flow direction. At higher viscosity ratios, a tumbling motion is observed in which the whole vesicle rotates with a periodically modulated velocity. When the shear rate increases, this tumbling motion becomes increasingly sensitive to vesicle deformation due to the elongational component of the flow and significant deviations from simpler models are observed. A good characterization of these various flow regimes is essential for the validation of analytical and numerical models, and to relate microscopic dynamics to macroscopic rheology of suspensions of deformable particles, such as blood.     

流场环境下复杂囊泡的动力学行为

采用非平衡态分子动力学方法研究了二维复杂囊泡在剪切流中的动力学行为. 模拟发现了复杂囊泡经典的翻滚(tumbling)、摇摆(trembling)和坦克履(tank-treading)行为, 还观察到由坦克履行为向平动行为(translating)的转变. 囊泡的平动行为与剪切率大小、复杂囊泡的形状密切相关. 当大囊泡均匀嫁接较多数目的小囊泡后, 其平动方式消失. 该研究有益于加深对囊泡在剪切流场中复杂性行为的理解.     

Dynamics and statics of DNA-programmable nanoparticle self-assembly and crystallization

DNA linker mediated self-assembly is emerging as a very general strategy for designing new materials. In this Letter, we characterize both the dynamics and thermodynamics of nanoparticle-DNA self-assembly by molecular dynamics simulations from a new coarse-grained model. We establish the general phase diagram and discuss the stability of a previously overlooked crystalline phase (D-bcc). We also characterize universal properties about the dynamics of crystallization. We point out the connection to f-star polymer systems and discuss the implications for ongoing experiments as well as for the general field of DNA mediated self-assembly.     

Dynamics of multicomponent vesicles in a viscous fluid

We develop and investigate numerically a thermodynamically consistent model of two-dimensional multicomponent vesicles in an incompressible viscous fluid. The model is derived using an energy variation approach that accounts for different lipid surface phases, the excess energy (line energy) associated with surface phase domain boundaries, bending energy, spontaneous curvature, local inextensibility and fluid flow via the Stokes equations. The equations are high-order (fourth order) nonlinear and nonlocal due to incompressibility of the fluid and the local inextensibility of the vesicle membrane. To solve the equations numerically, we develop a nonstiff, pseudo-spectral boundary integral method that relies on an analysis of the equations at small scales. The algorithm is closely related to that developed very recently by Veerapaneni et al. [81] for homogeneous vesicles although we use a different and more efficient time stepping algorithm and a reformulation of the inextensibility equation. We present simulations of multicomponent vesicles in an initially quiescent fluid and investigate the effect of varying the average surface concentration of an initially unstable mixture of lipid phases. The phases then redistribute and alter the morphology of the vesicle and its dynamics. When an applied shear is introduced, an initially elliptical vesicle tank-treads and attains a steady shape and surface phase distribution. A sufficiently elongated vesicle tumbles and the presence of different surface phases with different bending stiffnesses and spontaneous curvatures yields a complex evolution of the vesicle morphology as the vesicle bends in regions where the bending stiffness and spontaneous curvature are small.     

Dynamics of domain growth in self-assembled fluid vesicles

The dynamics of phase separation in multicomponent bilayer fluid vesicles is investigated by means of large-scale dissipative particle dynamics. The model explicitly accounts for solvent particles, thereby allowing for the very first numerical investigation of the effects of hydrodynamics and area-to-volume constraints. We observed regimes corresponding to coalescence of flat patches, budding and vesiculation, and coalescence of caps. We point out that the area-to-volume constraint has a strong influence on crossovers between these regimes.     

Influence of lipid composition and membrane curvature on fluorescence and solvent relaxation kinetics in unilamellar vesicles

Time-resolved fluorescence on unilamellar vesicles shows that increasing amounts of anionic, natural lipid lead to a larger increase in polarity close to the headgroups than in the hydrophobic core of the bilayer. The region close to the headgroups is less polar in vesicles containing phosphatic acid rather than phosphatidylserine. A greater membrane curvature increases the mobility of the hydrated headgroups.     

Effect of C_(60) nanoparticles on elasticity of small unilamellar vesicles composed of DPPC bilayers

The interaction between C60 nanoparticles and biomembranes has been of great interest in researches over the past decades due to their novel applications as well as potential cytotoxicity.In this work,we study the deformation of the small unilamellar vesicles composed of dipalmitoylphosphatidylcholine(DPPC)lipid bilayers infiltrated with C60 nanoparticles of different molecular concentrations through coarse-grained molecular dynamics simulations.By employing the Helfrich spontaneous curvature model,the bending modulus and the spontaneous curvature of the vesicles with C60 nanoparticles of different concentrations are obtained according to the simulation data.The results show that the bending modulus and the spontaneous curvature of pure DPPC vesicle membranes are approximately 1.6×10^-19J and 1.4 nm^-1,respectively.Both of them increase linearly approximately as the C60 concentration increases from 0 to 16.3%.The density profiles of vesicles,the order of lipid packing and the diffusion characteristics of DPPC and C60 are also investigated.