Enhanced chromosome extraction from cells using a pinched flow microfluidic device.

Enhanced chromosome extraction from cells using a pinched flow microfluidic device. Biomed Microdevices. 2020 Mar 12;22(2):25 Authors: Feng H, Hockin M, Zhang S, Capecchi M, Gale B, Sant H Abstract Extraction and purification of intact chromosomes are critical sample preparation steps for transchromosomic research and other applications. The commonly used sample preparation methods lead to too few chromosomes with chromosome deactivation and degradation. In this paper, a "mild" chromosome extraction process that combines a chemical and mechanical lysis approach is introduced for the preparation of intact chromosomes that can readily be used for downstream processing. Metaphase cells are treated by chemical lysis buffer and pushed through a microfluidic pinched flow device. Cells are ruptured, and chromosomes are released by a combination of shear stress and chemical reagents. Chromosomes are released intact from the cell membrane into the solution. Simulations and experiments are performed to optimize the microfluidic device geometry and operation parameters. Cell rupture and chromosome release are found to be improved by the shear stress in the pinched flow device. Simulation results indicate that the maximum shear stress appears in the channel constriction region, and the narrow channel maintains constant shear stress. It is concluded that the constriction design, narrow channel width, and operation flow rate have a significate inf...
Source: Biomedical Microdevices - Category: Biomedical Engineering Authors: Tags: Biomed Microdevices Source Type: research