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Single-lipid dynamics in phase-separated supported lipid bilayers
Chemistry and Physics of Lipids ( IF 3.4 ) Pub Date : 2020-10-26 , DOI: 10.1016/j.chemphyslip.2020.104991
Xinxin Woodward 1 , Christopher V Kelly 1
Affiliation  

Phase separation is a fundamental organizing mechanism on cellular membranes. Lipid phases have complex dependencies on the membrane composition, curvature, tension, and temperature. Lipid diffusion rates vary by up to ten-fold between liquid-disordered (Ld) and liquid-ordered (Lo) phases depending on the membrane composition, measurement technique, and the surrounding environment. This manuscript reports the lipid diffusion on phase-separated supported lipid bilayers (SLBs) with varying temperature, composition, and lipid phase. Lipid diffusion is measured by single-particle tracking (SPT) and fluorescence correlation spectroscopy (FCS) via custom data acquisition and analysis protocols that apply to diverse membranes systems. Traditionally, SPT is sensitive to diffuser aggregation, whereas the diffusion rates reported by FCS are unaffected by the presence of immobile aggregates. Within this manuscript, we report (1) improved single-particle tracking analysis of lipid diffusion, (2) comparison and consistency between diffusion measurement methods for non-Brownian diffusers, and (3) the application of these methods to measure the phase, temperature, and composition dependencies in lipid diffusion. We demonstrate improved SPT analysis methods that yield consistent FCS and SPT diffusion results even when most fluorescent lipids are frequently confined within aggregates within the membrane. With varying membrane composition and temperature, we demonstrate differences in diffusion between the Ld and Lo phases of SLBs.



中文翻译:

相分离的支持脂质双层中的单脂质动力学

相分离是细胞膜上的基本组织机制。脂质相对膜成分、曲率、张力和温度具有复杂的依赖性。液体无序 ( L d ) 和液体有序 ( L o) 相取决于膜成分、测量技术和周围环境。这份手稿报告了具有不同温度、组成和脂质相的相分离支撑脂质双层 (SLB) 上的脂质扩散。通过适用于不同膜系统的自定义数据采集和分析协议,通过单粒子跟踪 (SPT) 和荧光相关光谱 (FCS) 测量脂质扩散。传统上,SPT 对扩散器聚集很敏感,而 FCS 报告的扩散速率不受固定聚集体存在的影响。在本手稿中,我们报告了 (1) 改进的脂质扩散单粒子跟踪分析,(2) 非布朗扩散器扩散测量方法之间的比较和一致性,(3) 应用这些方法测量脂质扩散中的相、温度和成分依赖性。我们展示了改进的 SPT 分析方法,即使大多数荧光脂质经常被限制在膜内的聚集体中,也能产生一致的 FCS 和 SPT 扩散结果。随着膜组成和温度的变化,我们证明了膜之间的扩散差异SLB 的L dL o相。

更新日期:2020-11-03
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