Yang Anxiong, Lein Falk Nicolas, Weiler Joana, Drechsel Julian, Schumann Vanessa, Erichson Felix, Streek André, Börner Richard
Laserinstitut Hochschule Mittweida, University of Applied Science Mittweida, Technikumplatz 17, 09648 Mittweida, Germany.
HardwareX. 2023 Apr 27;14:e00425. doi: 10.1016/j.ohx.2023.e00425. eCollection 2023 Jun.
Sample preparation is a crucial step in single-molecule experiments and involves passivating the microfluidic sample chamber, immobilizing the molecules, and setting experimental buffer conditions. The efficiency of the experiment depends on the quality and speed of sample preparation, which is often performed manually and relies on the experience of the experimenter. This can result in inefficient use of single-molecule samples and time, especially for high-throughput applications. To address this, a pressure-controlled microfluidic system is proposed to automate single-molecule sample preparation. The hardware is based on microfluidic components from ElveFlow and is designed to be cost-effective and adaptable to various microscopy applications. The system includes a reservoir pressure adapter and a reservoir holder designed for additive manufacturing. Two flow chamber designs Ibidi µ-slide and Grace Bio-Labs HybriWell chamber are characterized, and the flow characteristics of the liquid at different volume flow rates are simulated using CFD-simulations and compared to experimental and theoretical values. The goal of this work is to establish a straightforward and robust system for single-molecule sample preparation that can increase the efficiency of experiments and reduce the bottleneck of manual sample preparation, particularly for high-throughput applications.
样品制备是单分子实验中的关键步骤，包括对微流控样品腔进行钝化、固定分子以及设置实验缓冲条件。实验效率取决于样品制备的质量和速度，而样品制备通常是手动进行的，并且依赖于实验者的经验。这可能导致单分子样品和时间的低效利用，特别是对于高通量应用而言。为了解决这个问题，提出了一种压力控制的微流控系统来实现单分子样品制备的自动化。该硬件基于来自ElveFlow的微流控组件，设计具有成本效益且适用于各种显微镜应用。该系统包括一个储液器压力适配器和一个专为增材制造设计的储液器支架。对两种流动腔设计（Ibidi μ-slide和Grace Bio-Labs HybriWell腔）进行了表征，并使用CFD模拟对不同体积流速下液体的流动特性进行了模拟，并与实验值和理论值进行了比较。这项工作的目标是建立一个简单而强大的单分子样品制备系统，该系统可以提高实验效率并减少手动样品制备的瓶颈，特别是对于高通量应用。
