Fabrication of a new all-in-one microfluidic dielectrophoresis integrated chip and living cell separation

Abstract

Summary Microfluidic dielectrophoresis (DEP) technology has been applied to many devices to perform label-free target cell separation. Cells separated by these devices are used in laboratories, mainly for medical research. The present study designed a microfluidic DEP device to fabricate a rapid and semiautomated cell separation system in conjunction with microscopy to enumerate the separated cells. With this device, we efficiently segregated bacterial cells from liquid products and enriched one cell type from two mixed eukaryotic cell types. The device eliminated sample pretreatment and established cell separation by all-in-one operation in a lab-on-chip, requiring only a small sample volume (0.5–1 mL) to enumerate the target cells and completing the entire separation process within 30 min. Such a rapid cell separation technique is in high demand by many researchers to promptly characterize the target cells.

Protocol of living cell separation using the microfluidic dielectrophoresis integrated chip

“… Lock Chip Device Lever to set the chip position.

Figure 1 Schematic design of the microfluidic dielectrophoresis chip The figure is from Oshiro et al. (2022) .

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“…The anticipated outcome of this protocol is to selectively collect the target cells in Outlet 1. When an AC voltage with a specific frequency is applied to the electrodes, the electrodes capture the target cells flowing from the hydrodynamic filtration (HDF) region (buffer exchange region of the chip) into the dielectrophoresis (DEP) region by dielectrophoretic force, and they are discharged to Outlet Port 1 ( Oshiro et al., 2022 ). The cells that do not respond to a specific frequency are not captured by the electrodes and are discharged to Outlet Port 2.…”

“…Such instructions, although trivial, are critical for achieving cell separation. For complete details on the use and execution of this protocol, please refer to Oshiro et al. (2022) .…”

“…For complete details on the use and execution of this protocol, please refer to Oshiro et al. (2022) .…”

Efficient Capture and Separation of Cancer Cells Using Hyaluronic Acid-Modified Magnetic Beads in a Microfluidic Chip

“…Recently, microfluidic chip technology has aroused great interest due to its low sample cost, high speed, good flexibility, high-throughput performance, and suitability for integrating multiple functional units for cell manipulation. − Various types of microfluidic chips have been designed based on optical trapping, , dielectrophoresis, − inertial focusing, , magnetic tweezers, or acoustophoresis. , Although each technique has its own limitations because of the heterogeneity of tumor cells, it performs well in the enrichment of CTCs under certain conditions. Among these techniques, magnetic separation is a promising tool for CTC enrichment due to its easy manipulation, high capture efficiency, and convenient coupling with immunocytochemistry and the polymerase chain reaction assay. − …”