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Vimentin networks at tunable ion-concentration in microfluidic drops

机译:波形蛋白网络在微流控滴中的离子浓度可调

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摘要

The structure and function of biological systems, for example, cells and proteins, depend strongly on their chemical environment. To investigate such dependence, we design a polydimethylsiloxane-based microfluidic device to encapsulate biological systems in picoliter-sized drops. The content of each individual drop is tuned in a defined manner. As a key feature of our method, the individual chemical composition is determined and related to the drop content. In our case, the drop content is imaged using microscopy methods, while the drops are immobilized to allow for long-time studies. As an application of our device, we study the influence of divalent ions on vimentin intermediate filament networks in a quantitative way by tuning the magnesium concentration from drop to drop. This way we are able to directly image the effect of magnesium on the fluorescently tagged protein in a few hundreds of drops. Our study shows that with increasing magnesium concentration in the drops, the compaction of the networks becomes more pronounced. The degree of compaction is characterized by different morphologies; freely fluctuating networks are observed at comparatively low magnesium concentrations of 5–10 mM, while with increasing magnesium concentration reaching 16 mM they develop into fully aggregated networks. Our approach demonstrates how a systematic study of interactions in biological systems can benefit from the exceptional controllability of microfluidic methods.
机译:生物系统(例如细胞和蛋白质)的结构和功能在很大程度上取决于其化学环境。为了研究这种依赖性,我们设计了一种基于聚二甲基硅氧烷的微流体装置,以将生物系统封装在皮升大小的液滴中。每个液滴的内容都以定义的方式进行调整。作为我们方法的关键特征,确定各个化学成分并与液滴含量相关。在我们的案例中,使用显微镜方法对液滴含量进行成像,同时固定液滴以进行长期研究。作为设备的一种应用,我们通过逐滴调整镁的浓度,定量研究了二价离子对波形蛋白中间丝网络的影响。这样,我们就可以在几百滴液滴中直接成像镁对荧光标记蛋白的作用。我们的研究表明,随着液滴中镁浓度的增加,网络的压缩变得更加明显。压实度的特征在于不同的形态。在相对较低的镁浓度为5-10 mM时观察到自由波动的网络,而随着镁浓度的增加达到16 mM时,它们发展为完全聚集的网络。我们的方法证明了如何系统地研究生物系统中的相互作用可以从微流体方法的出色可控性中受益。

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