George Annitta, Alva Ernesto, Brancaleon Lorenzo, Marucho Marcelo
Department of Physics and Astronomy, The University of Texas at San Antonio, San Antonio, Texas, United States of America.
PLoS One. 2024 Dec 31;19(12):e0312430. doi: 10.1371/journal.pone.0312430. eCollection 2024.
The accurate characterization of microtubules is essential for understanding their roles in various biological activities in eukaryotic cellular processes. In vitro, experimental data on these systems often need more details and information on sample preparation protocols and experimental techniques. This deficiency leads to unreproducible or contradictory outcomes. The use of diverse experimental methods and preparations yields different results of hydrodynamic and electro-mechanical properties, complicating the process of obtaining meaningful data and conclusive information. This article presents a robust and detailed protocol for performing dynamic light scattering (DLS) and electrophoretic light scattering (ELS) measurements on microtubules at low concentrations. This method ensures accurate and reproducible results on essential microtubule filament parameters such as the diffusion coefficient (D) and electrophoretic mobility (μ) from which other structures' hydrodynamics, electrical, and stability properties can be elucidated.
准确表征微管对于理解其在真核细胞过程中各种生物活动中的作用至关重要。在体外,关于这些系统的实验数据往往需要更多关于样品制备方案和实验技术的详细信息。这种不足导致结果不可重复或相互矛盾。使用不同的实验方法和制备方法会产生不同的流体动力学和机电性能结果,使得获取有意义的数据和确凿信息的过程变得复杂。本文介绍了一种稳健且详细的方案,用于在低浓度下对微管进行动态光散射(DLS)和电泳光散射(ELS)测量。该方法可确保在诸如扩散系数(D)和电泳迁移率(μ)等基本微管丝参数上获得准确且可重复的结果,从中可以阐明其他结构的流体动力学、电学和稳定性特性。