A continuous electrical cell lysis device using a low dc voltage for a cell transport and rupture

We present a continuous and low voltage cell lysis device in which a width and length of a channel change to generate focused the high electric field strength for cell lysis and the low electric field strength for a transport of samples. The previous cell lysis devices acquire the high electric field strength for a cell lysis by applying an ac voltage to a micro-gap between electrodes and require additional pumps or valves for a sample transport. However, when we change the width and length of the channel between a pair of external electrodes attached to a dc voltage, we generate both the high electric field strength for a cell lysis and the low electric field strength for an electroosmotic flow. The present device therefore performs continuous cell lysis and a sample transport without needing either an additional flow source or an additional process fabricating the electrodes for the micro-gap. The experimental study features an orifice whose width and length is 20 times narrower and 175 times shorter than the width and length of a microchannel. With an operational voltage of 50 V, the present device generates high electric field strength of 1.2 kV/cm at the orifice to disrupt cells with 100% lysis rate of red blood cells and low electric field strength of 60 V/cm at the microchannel to generate an electroosmotic flow of 30 ± 9 μm/s. In conclusion, the present device is capable of continuous self-pumping cell lysis at a low voltage; thus, it is suitable for a sample pretreatment component of a micro total analysis system or lab-on-a-chip.

[1]  Yong Huang,et al.  Microfabricated electroporation chip for single cell membrane permeabilization , 2001 .

[2]  Yu-Cheng Lin,et al.  Electroporation microchips for continuous gene transfection , 2001 .

[3]  Marc Madou,et al.  MEMS-based sample preparation for molecular diagnostics , 2002, Analytical and bioanalytical chemistry.

[4]  Zhao-Lun Fang,et al.  Integration of single cell injection, cell lysis, separation and detection of intracellular constituents on a microfluidic chip. , 2004, Lab on a chip.

[5]  K. Jensen,et al.  A microfluidic electroporation device for cell lysis. , 2005, Lab on a chip.

[6]  Y. Tai,et al.  A micro cell lysis device , 1999 .

[7]  Dino Di Carlo,et al.  On-chip cell lysis by local hydroxide generation. , 2005, Lab on a chip.

[8]  M. Heller,et al.  Electric manipulation of bioparticles and macromolecules on microfabricated electrodes. , 2001, Analytical chemistry.

[9]  Paul Yager,et al.  Cell lysis and protein extraction in a microfluidic device with detection by a fluorogenic enzyme assay. , 2002, Analytical chemistry.

[10]  Paul C. H. Li,et al.  Transport, manipulation, and reaction of biological cells on-chip using electrokinetic effects. , 1997, Analytical chemistry.

[11]  J. Graham,et al.  Subcellular fractionation : a practical approach , 1997 .

[12]  L J Bond,et al.  Continuous spore disruption using radially focused, high-frequency ultrasound. , 2001, Analytical chemistry.

[13]  U. Zimmermann,et al.  Electrical breakdown, electropermeabilization and electrofusion. , 1986, Reviews of physiology, biochemistry and pharmacology.