Journal article Open Access

An integrated microfluidic chip for chromosome enumeration using fluorescence in situ hybridization

Sieben, Vincent J.; Debes-Marun, Carina S.; Pilarski, Linda M.; Backhouse, Christopher J.

Fluorescence in-situ hybridization (FISH) is more sensitive than classical cytogenetics for detecting cryptic chromosomal abnormalities. H; however, the protocol complexity, high reagent cost and long hybridization time associated with a typical interphase FISH analysis have slowed its utilization in a clinical setting. For various cancers, such as multiple myeloma, the lack of a cost-effective and informative diagnostic method has compromised the quality of life for patients. Here, wWe present the first demonstration of a microchip-based FISH protocol for the analysis of chromosomal abnormalities in multiple myeloma cells to address this issue. The developed microfluidic arrays allow several chromosomal abnormalities associated with multiple myeloma to be detected with a 10 fold higher throughput and 1/10th the reagent consumption of the traditional slide based FISH. These benefits have resulted in the arrays being actively used in a clinical laboratory. We examined two methods of enhancing the hybridization, using mechanical and electrokinetic agitation. Both methods yielded improvements in the hybridization efficiency and warrant further optimization studies. Ultimately, we established a novel method of performing interphase FISH on a microchip in hours whereas the conventional protocol requires days. We believe that additional optimization studies would improve the hybridization enhancement even further, reducing the analysis time to less than one hour and making point of care FISH analysis a possibility.

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