Single-Molecule Diffusion and Assembly on Polymer-Crowded Lipid Membranes

Satyaghosh Maurya; Vishwesh Haricharan Rai; Aditya Upasani; Saurabh Umrao; Diksha Parwana; Rahul Roy

doi:10.3791/64243

JoVE Journal
Biochemistry
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JoVE Journal Biochemistry

Single-Molecule Diffusion and Assembly on Polymer-Crowded Lipid Membranes

聚合物拥挤脂质膜上的单分子扩散和组装

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10:43 min

July 19, 2022

DOI: 10.3791/64243-v

Satyaghosh Maurya¹, Vishwesh Haricharan Rai¹, Aditya Upasani¹, Saurabh Umrao^1,2, Diksha Parwana¹, Rahul Roy^1,3

¹Department of Chemical Engineering,Indian Institute of Science, ²Holonyak Micro & Nanotechnology Lab,University of Illinois, Urbana-Champaign, ³Center for BioSystems Science and Engineering,Indian Institute of Science

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在这里，提出了一种使用单分子全内反射荧光（smTIRF）显微镜执行和分析单分子在人工拥挤脂质膜上的结合，迁移率和组装的协议。

由于嵌入的蛋白质和糖的存在，细胞膜高度拥挤。我们在合成拥挤的脂质双层上演示了单分子成像。该平台可用于在分子水平上研究拥挤对膜生物分子反应的影响。

该协议解释了膜生物分子的结合，扩散和组装的分析。实验的成功取决于对底物的严格清洁，避免损坏双层，并在单分子成像中保持高信噪比。和我一起演示该程序的是Aditya Upasani和Vishwesh Haricharan Rai，他们是实验室的研究生。

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