A flow‐through microfluidic chip for continuous dielectrophoretic separation of viable and non‐viable human T‐cells

Abstract

Effective methods for rapid sorting of cells according to their viability are critical in T cells based therapies to prevent any risk to patients. In this context, we present a novel microfluidic device that continuously separates viable and non-viable T-cells according to their dielectric properties. A dielectrophoresis (DEP) force is generated by an array of castellated microelectrodes embedded into a microfluidic channel with a single inlet and two outlets; cells subjected to positive DEP forces are drawn toward the electrodes array and leave from the top outlet, those subjected to negative DEP forces are repelled away from the electrodes and leave from the bottom outlet. Computational fluid dynamics is used to predict the device separation efficacy, according to the applied alternative current (AC) frequency, at which the cells move from/to a negative/positive DEP region and the ionic strength of the suspension medium. The model is used to support the design of the operational conditions, confirming a separation efficiency, in terms of purity, of 96% under an applied AC frequency of 1.5 × 10 6 Hz and a flow rate of 20 μl/h. This work represents the first example of effective continuous sorting of viable and non-viable human T-cells in a single-inlet microfluidic chip, paving the way for lab-on-a-chip applications at the point of need.

Foundation of the Manipulation Technology for Tiny Objects Based on the Control of the Heterogeneity of Electric Fields

“…V was determined for each different position of the glass sphere by 0.5 mm. Subsequently, the net force exerted on the glass sphere at every position is determined by integrating the Maxwell stress tensor at sphere surface S, as described by Equation (9) [12]…”

“…If we develop a technique that can manipulate multiple target pieces individually and combine them with identification meanings (e.g., by visible and infrared light), most of the complexities involving sorting by physical properties can be removed. Applications of electrostatic forces other than plastic sorting include dust control on solar panels on the earth [2][3][4] and lunar surface [5], trapping plenty of particles onto insulator surfaces [6], sorting, trapping, traveling of living cells, and microparticles in the liquid phase [7][8][9][10][11][12][13][14][15][16][17][18][19]. Representative device structures are (a) a plane substrate with interdigitated electrodes on it [3][4][5][6]8,14,16,18], (b) a micro channel with multiple electrodes along it [7,9], and (c) a micro chamber surrounded by a set of electrodes on a plane [19].…”

“…Applications of electrostatic forces other than plastic sorting include dust control on solar panels on the earth [2][3][4] and lunar surface [5], trapping plenty of particles onto insulator surfaces [6], sorting, trapping, traveling of living cells, and microparticles in the liquid phase [7][8][9][10][11][12][13][14][15][16][17][18][19]. Representative device structures are (a) a plane substrate with interdigitated electrodes on it [3][4][5][6]8,14,16,18], (b) a micro channel with multiple electrodes along it [7,9], and (c) a micro chamber surrounded by a set of electrodes on a plane [19]. They apply phase-controlled AC voltage to treat target pieces not individually but as a mass.…”

Study on Microfluidic Chip Flow Rate Uniformity for Cell Activity Detection

“…With the introduction of micro-total-analysis systems and lab-on-a-chip, − cell viability detection tends to miniaturization, portability, low pollution, low cost, and easy operation. One of the most promising technologies is cell viability detection inside a microfluidic chip .…”

A novel single layer microfluidic device for dynamic stimulation, culture and imaging of mammalian cells

“…Microfluidic devices find their advantage in using low sample volumes, while allowing precise control over cell culture environment and mimicking physiological conditions [4][5][6][7] . Recently, there has been an increased interest to employ microfluidic devices for a range of mammalian cells applications, such as high throughput single cell screening [8][9][10] , parallelized drug screening 11 , temporal modulations of inputs 12 , easy-to-use perfusion for cell proliferation and differentiation 13,14 and automated feedback control of biological processes 15 .…”