Université Claude Bernard Lyon 1, CNRS, INSA Lyon, Ecole Centrale de Lyon, CPE Lyon, INL, Villeurbanne, France.
Université Claude Bernard Lyon 1, CNRS, Institut Lumière Matière, Villeurbanne, France.
Methods Mol Biol. 2024;2804:91-100. doi: 10.1007/978-1-0716-3850-7_5.
Circulating tumor cells (CTCs) isolated directly from whole blood opens new perspectives for cancer monitoring and the development of personalized treatments. However, due to their rarity among the multitude of blood cells, it remains a challenge to recover them alive with high level of purity, i.e., with few remaining white blood cells, and in a time frame compatible with the clinical context. Microfluidic chips have emerged as promising tools to address these challenges. We propose a two-step workflow including a pre-enrichment step, performed by a size-based pre-enrichment system, and a purification step, performed by an immunomagnetic chip. Here, we describe the protocol for the fabrication of the immunomagnetic microchip, the preparation of the sample, and the procedure for injection into the microchip allowing the sorting of the CTCs.
从全血中直接分离出的循环肿瘤细胞 (CTCs) 为癌症监测和个性化治疗的发展开辟了新的视角。然而，由于其在众多血细胞中非常罕见，因此仍然难以以高纯度（即白细胞残留少）和与临床环境兼容的时间框架将其回收。微流控芯片已成为解决这些挑战的有前途的工具。我们提出了一个两步工作流程，包括一个基于大小的预富集系统进行的预富集步骤和一个免疫磁珠芯片进行的纯化步骤。在这里，我们描述了免疫磁珠微芯片的制造协议、样品的准备以及将样品注入微芯片的程序，从而实现 CTC 的分选。
