Bhatia Tripta, Husen Peter, Ipsen John H, Bagatolli Luis A, Simonsen Adam Cohen
MEMPHYS - Center for Biomembrane Physics, University of Southern Denmark (SDU), 5230 Odense M, Denmark; Department of Physics Chemistry and Pharmacy, SDU.
MEMPHYS - Center for Biomembrane Physics, University of Southern Denmark (SDU), 5230 Odense M, Denmark.
Biochim Biophys Acta. 2014 Oct;1838(10):2503-10. doi: 10.1016/j.bbamem.2014.05.016. Epub 2014 May 24.
We devise a methodology to fixate and image dynamic fluid domain patterns of giant unilamellar vesicles (GUVs) at sub-optical length scales. Individual GUVs are rapidly transferred to a solid support forming planar bilayer patches. These are taken to represent a fixated state of the free standing membrane, where lateral domain structures are kinetically trapped. High-resolution images of domain patterns in the liquid-ordered (lo) and liquid-disordered (ld) co-existence region in the phase-diagram of ternary lipid mixtures are revealed by atomic force microscopy (AFM) scans of the patches. Macroscopic phase separation as known from fluorescence images is found, but with superimposed fluctuations in the form of nanoscale domains of the lo and ld phases. The size of the fluctuating domains increases as the composition approaches the critical point, but with the enhanced spatial resolution, such fluctuations are detected even deep in the coexistence region. Agreement between the area-fraction of domains in GUVs and the patches respectively, supports the assumption that the thermodynamic state of the membrane remains stable. The approach is not limited to specific lipid compositions, but could potentially help uncover lateral structures in highly complex membranes.
我们设计了一种方法,用于在亚光学长度尺度下固定和成像巨型单层囊泡(GUVs)的动态流体域模式。将单个GUVs快速转移到固体支持物上,形成平面双层斑块。这些斑块被视为代表自由站立膜的固定状态,其中横向域结构在动力学上被捕获。通过对斑块进行原子力显微镜(AFM)扫描,揭示了三元脂质混合物相图中液相有序(lo)和液相无序(ld)共存区域的域模式的高分辨率图像。发现了荧光图像中已知的宏观相分离,但伴有lo相和ld相纳米级域形式的叠加波动。随着组成接近临界点,波动域的尺寸增大,但由于空间分辨率提高,即使在共存区域深处也能检测到这种波动。GUVs和斑块中域的面积分数之间的一致性,支持了膜的热力学状态保持稳定的假设。该方法不限于特定的脂质组成,而是有可能帮助揭示高度复杂膜中的横向结构。