Francis Crick Institute, London, United Kingdom.
PLoS One. 2020 Dec 15;15(12):e0244103. doi: 10.1371/journal.pone.0244103. eCollection 2020.
We present a fast, inexpensive and robust technique for constructing thin, optically transparent flow-cells with pump-free flow control. Using layers of glass, patterned adhesive tape and polydimethylsiloxane (PDMS) connections, we demonstrate the fabrication of planar devices with chamber height as low as 25 μm and with millimetre-scale (x,y) dimensions for wide-field microscope observation. The method relies on simple benchtop equipment and does not require microfabrication facilities, glass drilling or other workshop infrastructure. We also describe a gravity perfusion system that exploits the strong capillary action in the flow chamber as a passive limit-valve. Our approach allows simple sequential sample exchange with controlled flow rates, sub-5 μL sample chamber size and zero dead volume. We demonstrate the system in a single-molecule force spectroscopy experiment using magnetic tweezers.
我们提出了一种快速、廉价且稳健的技术，用于构建带有无泵流量控制的薄型、光学透明的流动池。使用玻璃层、图案化的压敏胶带和聚二甲基硅氧烷 (PDMS) 连接，我们展示了具有低至 25 μm 腔室高度的平面器件的制造方法，以及具有毫米级 (x,y) 尺寸的平面器件，可用于宽场显微镜观察。该方法依赖于简单的台式设备，不需要微制造设施、玻璃钻孔或其他车间基础设施。我们还描述了一种重力灌注系统，该系统利用流动池中的强毛细作用作为被动限流器。我们的方法允许简单的顺序样品交换，具有可控的流速、亚 5 μL 的样品腔尺寸和零死体积。我们在使用磁镊的单分子力谱实验中展示了该系统。
