Sieben V J, Debes Marun C S, Pilarski P M, Kaigala G V, Pilarski L M, Backhouse C J
University of Alberta, Department of Electrical and Computer Engineering, CanadaCross Cancer Institute, Edmonton, Canada.
IET Nanobiotechnol. 2007 Jun;1(3):27-35. doi: 10.1049/iet-nbt:20060021.
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.
间期荧光原位杂交(FISH)是一种灵敏的诊断工具,用于逐个细胞检测基因组的改变。然而,当前FISH检测方法的每次检测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,缺乏一种经济高效且信息丰富的诊断方法影响了患者的生活质量。我们首次展示了一种基于微芯片的FISH检测方法,并结合了一种将外周血单个核细胞固定在微流控通道内的新方法。这些首次在芯片上实现的FISH检测能够检测与多发性骨髓瘤相关的几种染色体异常,通量比传统基于玻片的方法高十倍,试剂消耗仅为其十分之一。此外,芯片检测在数小时内即可完成,而传统检测方法则需要数天时间。另外,还研究了两种用于增强FISH杂交效果的芯片上方法:机械泵和电动泵。类似的搅拌方法在DNA微阵列工作中显著提高了杂交效率,但对于这种基于细胞的方法,效果较为适中。芯片上FISH技术有望在每次门诊时为癌症患者提供精密且经济高效的筛查。