Perez-Toralla Karla, Mottet Guillaume, Tulukcuoglu-Guneri Ezgi, Champ Jérôme, Bidard François-Clément, Pierga Jean-Yves, Klijanienko Jerzy, Draskovic Irena, Malaquin Laurent, Viovy Jean-Louis, Descroix Stéphanie
Macromolecules and Microsystems in Biology and Medicine, Institut Curie, Centre National de la Recherche Scientifique, Université Pierre et Marie Curie, PSL Research University, UMR 168, 75005, Paris, France.
Univ Paris Diderot, Sorbonne Paris Cité, F-75205, Paris, France.
Methods Mol Biol. 2017;1547:211-220. doi: 10.1007/978-1-4939-6734-6_16.
Microfluidics offer powerful tools for the control, manipulation, and analysis of cells, in particular for the assessment of cell malignancy or the study of cell subpopulations. However, implementing complex biological protocols on chip remains a challenge. Sample preparation is often performed off chip using multiple manually performed steps, and protocols usually include different dehydration and drying steps that are not always compatible with a microfluidic format.Here, we report the implementation of a Fluorescence in situ Hybridization (FISH) protocol for the molecular typing of cancer cells in a simple and low-cost device. The geometry of the chip allows integrating the sample preparation steps to efficiently assess the genomic content of individual cells using a minute amount of sample. The FISH protocol can be fully automated, thus enabling its use in routine clinical practice.
微流控技术为细胞的控制、操作和分析提供了强大的工具,特别是在评估细胞恶性程度或研究细胞亚群方面。然而,在芯片上实施复杂的生物学实验方案仍然是一项挑战。样品制备通常在芯片外通过多个手动步骤进行,而且实验方案通常包括不同的脱水和干燥步骤,这些步骤并不总是与微流控形式兼容。在此,我们报告了在一种简单且低成本的设备中实施用于癌细胞分子分型的荧光原位杂交(FISH)实验方案。芯片的几何结构允许整合样品制备步骤,从而能够使用微量样品有效地评估单个细胞的基因组内容。FISH实验方案可以完全自动化,因此能够在常规临床实践中使用。