Institut Curie, Laboratoire Physico-Chimie (CNRS UMR 168), Institut Pierre-Gilles de Gennes, Sorbonne Université, PSL Research University, Paris, France.
Institut Curie, UMR 168, Institut Pierre-Gilles de Gennes, Paris, France.
Methods Mol Biol. 2024;2804:163-176. doi: 10.1007/978-1-0716-3850-7_10.
Multiomics studies at single-cell level require small volume manipulation, high throughput analysis, and multiplexed detection, characteristics that droplet microfluidics can tackle. However, the initial step of molecule bioseparation remains challenging. Here, we describe a unique magnetic device to trap and extract magnetic particles in sub-nanoliter droplets, for compartmentalisation of detection steps. Relying on electrodeposition of NiFe structures and microfluidic manipulation, the extraction of 1 μm diameter magnetic particles was achieved at high throughput (20 droplets per second) with an efficiency close to 100% in 450 pL droplets. The first demonstration of its adaptability to single-cell analysis is demonstrated with the extraction of mRNA. Using a purified nucleic acid solution, this unique magnetic configuration was able to reach a RNA extraction rate of 72%. This is the first demonstration of a physical separation in droplets at high throughput at single-cell scale.
单细胞水平的多组学研究需要小体积操作、高通量分析和多重检测，这些都是液滴微流控技术可以解决的问题。然而，分子生物分离的初始步骤仍然具有挑战性。在这里，我们描述了一种独特的磁性装置，可以在亚纳升级别的液滴中捕获和提取磁性颗粒，用于分隔检测步骤。该装置依赖于 NiFe 结构的电沉积和微流控操作，以 20 个/秒的高通量在 450pL 液滴中实现了 1μm 直径磁性颗粒的高效提取，效率接近 100%。通过提取 mRNA ，首次展示了其在单细胞分析中的适应性。使用纯化的核酸溶液，这种独特的磁性结构能够达到 72%的 RNA 提取率。这是在单细胞尺度的高通量液滴中进行物理分离的首次演示。
