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质量敏感颗粒跟踪，用于表征膜相关的高分子动力学

JoVE Journal
Biochimica

This content is Free Access.

JoVE Journal Biochimica

Mass-Sensitive Particle Tracking to Characterize Membrane-Associated Macromolecule Dynamics

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质量敏感颗粒跟踪，用于表征膜相关的高分子动力学

DOI:

10.3791/63583-v

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13:30 min

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February 18, 2022

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Frederik Steiert², Tamara Heermann, Nikolas Hundt, Petra Schwille

¹Department of Cellular and Molecular Biophysics,Max Planck Institute of Biochemistry, ²Department of Physics,Technical University Munich, ³Department of Cellular Physiology, Biomedical Center (BMC),Ludwig-Maximilians-Universität München

Capitoli

00:05Introduction
00:43Cleaning of Microscope Slides
01:48Assembly of Flow Chambers
02:41Supported Lipid Bilayer (SLB) Formation
03:31Imaging
04:31Data Analysis
10:25Results: Native and Ratiometric View of Exemplary Surfaces
11:01Results: Calibration of the Mass-to-Contrast Relation for MSPT Measurements
12:15Results: Deciphering Oligomer States of Membrane-Associated Proteins
12:57Conclusion

Summary

Traduzione automatica

该协议描述了一种基于iSCAT的图像处理和单粒子跟踪方法，可以同时研究与脂质膜相互作用的大分子的分子质量和扩散行为。在提供样品制备、质量到对比度转换、电影采集和后处理的分步说明的同时，还提供了相应的说明，以防止潜在的陷阱。

Tags

Mass-sensitive Particle Tracking Membrane-associated Macromolecules Label-free Oligomeric States Lipid Membranes Biomolecule Dynamics Ligand-induced Receptor Complexes Small Unilamellar Vesicles Flow Chamber Mass Photometer

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