We present a novel label-free passive microfluidic technique for isolation of cancer cells (EpCAM+ and CD45−) from peripheral blood mononuclear cells (PBMCs) (CD45+ and EpCAM−) in aqueous two-phase system (ATPS).

Cell lysis is a critical step in genomics for the extraction of cellular components of downstream assays. Electrical lysis (EL) offers key advantages in terms of speed and non-interference. Here, we report a simple, chemical-free, and automated technique based on a microfluidic device with passivated interdigitated electrodes with DC fields for continuous EL of cancer cells. We show that the critical problems in EL, bubble formation and electrode erosion that occur at high electric fields, can be circumvented by passivating the electrodes with a thin layer (∼18 μm) of polydimethylsiloxane. We present a numerical model for the prediction of the transmembrane potential (TMP) at different coating thicknesses and voltages to verify the critical TMP criterion for EL. Our simulations showed that the passivation layer results in a uniform electric field in the electrode region and offers a TMP in the range of 5-7 V at an applied voltage of 800 V, which is well above the critical TMP (∼1 V) required for EL. Experiments revealed that lysis efficiency increases with an increase in the electric field (E) and residence time (t r ): a minimum E ∼ 10 5 V/m and t r ∼ 1.0 s are required for efficient lysis. EL of cancer cells is demonstrated and characterized using immunochemical staining and compared with chemical lysis. The lysis efficiency is found to be ∼98% at E = 4 × 10 5 V/m and t r = 0.72 s. The efficient recovery of genomic DNA via EL is demonstrated using agarose gel electrophoresis, proving the suitability of our method for integration with downstream on-chip assays.

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