A microfluidic flow-through device for high throughput electrical lysis of bacterial cells based on continuous dc voltage.
Interest in electrical lysis of biological cells on a microfludic platform has increased because it allows for the rapid recovery of intracellular contents without introducing lytic agents. In this study we demonstrated a simple microfluidic flow-through device which lysed Escherichia coli cells under a continuous dc voltage. The E. coli cells had previously been modified to express green fluorescent protein (GFP). In our design, the cell lysis only happened in a defined section of a microfluidic channel due to the local field amplification by geometric modification. The geometric modification also effectively decreased the required voltage for lysis by several folds. We found that local field strength of 1000-1500 V/cm was required for nearly 100% cell death. This threshold field strength was considerably lower than the value reported in the literature, possibly due to the longer duration of the field [Lee, S.W., Tai, Y.C., 1999. Sens. Actuators A: Phys. 73, 74-79]. Cell lysis was detected by both plate count and fluorescence spectroscopy. The cell membrane was completely disintegrated in the lysis section of the microfluidic device, when the field strength was higher than 2000 V/cm. The devices were fabricated using low-cost soft lithography with channel widths considerably larger than the cell size to avoid clogging and ensure stable performance. Our tool will be ideal for high throughput processing of bacterial cells for chemical analysis of intracellular contents such as DNA and proteins. The application of continuous dc voltage greatly simplified the instrumentation compared to devices using electrical pulses for similar purposes. In principle, the same approach can also be applied for lysis of mammalian cells and electroporative transfection.
Wang
Hsiang-Yu
HY
School of Chemical Engineering, Purdue University, West Lafayette, IN 47907, USA.
Bhunia
Arun K
AK
Lu
Chang
C
D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D002457:Cell Extracts / Q000302:isolation & purification*; D002458:Cell Fractionation / Q000295:instrumentation* / Q000379:methods; D004574:Electromagnetic Fields; D018274:Electroporation / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis; D004926:Escherichia coli / Q000166:cytology / Q000502:physiology* / Q000528:radiation effects; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods
D002457:Cell Extracts