Dynamic monitoring of single cell lysis in an impedance-based microfluidic device.
A microfluidic device that is capable of trapping and sensing dynamic variations in the electrical properties of individual cells is demonstrated. The device is applied to the real-time recording of impedance measurements of mouse embryonic stem cells (mESCs) during the process of membrane lysis, with the resulting changes in the electrical properties of cells during this process being quantitatively tracked over time. It is observed that the impedance magnitude decreases dramatically after cell membrane lysis. A significant shift in the phase spectrum is also observed during the time course of this process. By fitting experimental data to physical models, the electrical parameters of cells can be extracted and parameter variations quantified during the process. In the cell lysis experiments, the equivalent conductivity of the cell membrane is found to increase significantly due to pore formation in the membrane during lysis. An increase in the specific capacitance of the membrane is also observed. On the other hand, the conductivity of the cytoplasm is observed to decrease, which may be explained the fact that excess water enters the cell through the gradual permeabilization of the membrane during lysis. Cells can be trapped in the device for periods up to several days, and their electrical response can be monitored by real-time impedance measurements in a label-free and non-invasive manner. Furthermore, due to the highly efficient single cell trapping capacity of the device, a number of cells can be trapped and held in separate wells for concurrent parallel experiments, allowing for the possibility of stepped parametric experiments and studying cell heterogeneity by combining measurements across the array.
Department of Engineering, Nanoscience Centre, University of Cambridge, 11 JJ Thomson Avenue, Cambridge, CB3 0FF, UK.
Ying
Y
Zhou
Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge, CB2 1GA, UK.
Srinjan
S
Basu
Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge, CB2 1GA, UK.
Ernest D
ED
Laue
Department of Engineering, Nanoscience Centre, University of Cambridge, 11 JJ Thomson Avenue, Cambridge, CB3 0FF, UK. aas41@cam.ac.uk.
Ashwin A
AA
Seshia
D000818:Animals; D002462:Cell Membrane; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D025802:Electric Capacitance; D017097:Electric Impedance; D053595:Embryonic Stem Cells / Q000166:cytology; D004867:Equipment Design; D005069:Evaluation Studies as Topic; D056656:Lab-On-A-Chip Devices; D051379:Mice; D046210:Microfluidic Analytical Techniques; D008962:Models, Theoretical; D059010:Single-Cell Analysis
D004129:Dimethylpolysiloxanes; C013830:baysilon
Impedance spectroscopy; Microfluidics; Real-time measurements; Single cell trapping; Stem cells