FISH and chips: chromosomal analysis on microfluidic platforms
FISH and chips: chromosomal analysis on microfluidic platforms
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DOI：
10.1049/iet-nbt:20060021
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发表时间：
2007-06-01
影响因子：
2.3
通讯作者：
Backhouse, C. J.
中科院分区：
文献类型：
--
作者：
Sieben, V. J.;Marun, C. S. Debes;Backhouse, C. J.
间期荧光原位杂交（FISH）是一种灵敏的诊断工具，用于检测细胞基因组的变化。然而，当前FISH方案的每次测试成本和技术复杂性阻碍了其在临床环境中的广泛应用。对于许多癌症来说，缺乏具有成本效益和信息丰富的诊断方法已经损害了患者的生活质量。我们首次展示了一种基于微芯片的FISH方案，以及一种将外周血单个核细胞固定在微流体通道内的新方法。这些首次在芯片上实现的FISH允许检测几种与多发性骨髓瘤相关的染色体异常，其吞吐量提高了10倍，试剂消耗仅为传统基于载玻片的方法的1/10。此外，芯片测试可以在数小时内完成，而传统方案需要数天。此外，还研究了两种增强FISH杂交方面的芯片上方法：机械泵送和电动泵送。类似的搅拌方法已经显著提高了DNA微阵列工作的杂交效率，但这种基于细胞的方法的好处是中等的。芯片上的FISH技术有望在每次就诊时对癌症患者进行复杂且具有成本效益的筛查。
Interphase fluorescence in situ hybridisation (FISH) is a sensitive diagnostic tool used for the detection of alterations in the genome on cell-by-cell basis. However, the cost-per-test and the technical complexity of current FISH protocols have slowed its widespread utilisation in clinical settings. For many cancers, the lack of a cost-effective and informative diagnostic method has compromised the quality of life for patients. We present the first demonstration of a microchip-based FISH protocol, coupled with a novel method to immobilise peripheral blood mononuclear cells inside microfluidic channels. These first on-chip implementations of FISH allow several chromosomal abnormalities associated with multiple myeloma to be detected with a ten-fold higher throughput and 1/10-th the reagent consumption of the traditional slide-based method. Moreover, the chip test is performed within hours whereas the conventional protocol required days. In addition, two on-chip methods to enhance the hybridisation aspects of FISH have been examined: mechanical and electrokinetic pumping. Similar agitation methods have led to significant improvements in hybridisation efficiency with DNA microarray work, but with this cell-based method the benefits were moderate. On-chip FISH technology holds promise for sophisticated and cost-effective screening of cancer patients at every clinic visit.