Lycans Rebecca M, Higgins Catherine B, Tanner Michael S, Blough Eric R, Day B Scott
Department of Chemistry, Marshall University, Huntington, WV 25755, United States.
School of Pharmacy, Marshall University, Huntington, WV 25755, United States; Center for Diagnostic Nanosystems, Marshall University, Huntington, WV 25755, United States.
Colloids Surf B Biointerfaces. 2014 Apr 1;116:687-94. doi: 10.1016/j.colsurfb.2013.11.007. Epub 2013 Nov 15.
In vitro motility assays are readily used to simplify the complex environments within the cell and in muscle tissue. These assays have afforded considerable insight into the fundamentals of their underlying biophysics, interactions with cargo, intracellular regulation, and motor cooperation/competition. Extension of the standard in vitro motility assay into a more automated and cost-effective fluidic design while providing availability to the scientific community without expertise in lithographic fabrication is critical for the continued advancement of the field. In this work, we utilized a standard plasma cleaner to oxidize the widely prevalent material polydimethylsiloxane (PDMS) to create flow cells that could be used for in vitro motility assays. Our analysis indicated that a 40 min pre-treatment of the PDMS with plasma exposure resulted in optimal bundle motility. This finding was attributed to the condition at which the least amount of oxygen permeates the PDMS slab, enters the motility buffer, and oxidizes the motor proteins. Based on these findings, we developed a method for constructing microfluidic devices from glass and plasma-treated PDMS molds in which motility could be observed.
体外运动分析易于用于简化细胞和肌肉组织内的复杂环境。这些分析为其潜在生物物理学的基本原理、与货物的相互作用、细胞内调节以及马达协作/竞争提供了相当多的见解。将标准体外运动分析扩展为更自动化且成本效益更高的流体设计,同时为没有光刻制造专业知识的科学界提供可用性,对于该领域的持续发展至关重要。在这项工作中,我们利用标准等离子体清洗器氧化广泛存在的材料聚二甲基硅氧烷(PDMS),以创建可用于体外运动分析的流动池。我们的分析表明,用等离子体暴露对PDMS进行40分钟的预处理可产生最佳的束状运动。这一发现归因于氧气渗透到PDMS平板、进入运动缓冲液并氧化运动蛋白的量最少的条件。基于这些发现,我们开发了一种用玻璃和经等离子体处理的PDMS模具构建微流控装置的方法,在该装置中可以观察到运动。