化学
膜
生物物理学
化学物理
扩散
脂质双层
分子
跟踪(教育)
分子动力学
相(物质)
细胞膜
生物系统
纳米技术
生物化学
材料科学
计算化学
热力学
物理
心理学
教育学
有机化学
生物
作者
Jiseong Park,Yongdeok Ahn,Wonhee John Lee,Siwoo Jin,Sejoo Jeong,Jae-Yong Kim,Young‐Sam Lee,Jong Chan Lee,Daeha Seo
标识
DOI:10.1021/acs.analchem.3c02655
摘要
In live cells, the plasma membrane is composed of lipid domains separated by hundreds of nanometers in dynamic equilibrium. Lipid phase separation regulates the trafficking and spatiotemporal organization of membrane molecules that promote signal transduction. However, visualizing domains with adequate spatiotemporal accuracy remains challenging because of their subdiffraction limit size and highly dynamic properties. Here, we present a single lipid-molecular motion analysis pipeline (lipid-MAP) for analyzing the phase heterogeneity of lipid membranes by detecting the instantaneous velocity change of a single lipid molecule using the excellent optical properties of nanoparticles, high spatial localization accuracy of single-molecule localization microscopy, and separation capability of the diffusion state of the hidden Markov model algorithm. Using lipid-MAP, individual lipid molecules were found to be in dynamic equilibrium between two statistically distinguishable phases, leading to the formation of small (∼170 nm), viscous (2.5× more viscous than surrounding areas), and transient domains in live cells. Moreover, our findings provide an understanding of how membrane compositional changes, i.e., cholesterol and phospholipids, affect domain formation. This imaging method can contribute to an improved understanding of spatiotemporal-controlled membrane dynamics at the molecular level.
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