Impedance spectroscopy flow cytometry: on-chip label-free cell differentiation.
BACKGROUND
The microfabricated impedance spectroscopy flow cytometer used in this study permits rapid dielectric characterization of a cell population with a simple microfluidic channel. Impedance measurements over a wide frequency range provide information on cell size, membrane capacitance, and cytoplasm conductivity as a function of frequency. The amplitude, opacity, and phase information can be used for discrimination between different cell populations without the use of cell markers.


METHODS
Polystyrene beads, red blood cells (RBCs), ghosts, and RBCs fixed in glutaraldehyde were passed through a microfabricated flow cytometer and measured individually by using two simultaneously applied discrete frequencies. The cells were characterized at 1,000 per minute in the frequency range of 350 kHz to 20 MHz.


RESULTS
Cell size was easily measured with submicron accuracy. Polystyrene beads and RBCs were differentiated using opacity. RBCs and ghosts were differentiated using phase information, whereas RBCs and fixed RBCs were differentiated using opacity. RBCs fixed using increasing concentrations of glutaraldehyde showed increasing opacity. This increased opacity was linked to decreased cytoplasm conductivity and decreased membrane capacitance, both resulting from protein cross-linking.


CONCLUSIONS
This work presents label-free differentiation of cells in an on-chip flow cytometer based on impedance spectroscopy, which will be a powerful tool for cell characterization.
Cheung
Karen
K
Institute of Microelectronics and Microsystems, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland. karen.cheung@epfl.ch
Gawad
Shady
S
Renaud
Philippe
P
D002454:Cell Differentiation; D004553:Electric Conductivity; D017097:Electric Impedance; D004566:Electrodes; D004910:Erythrocyte Membrane / Q000378:metabolism / Q000502:physiology; D004912:Erythrocytes / Q000166:cytology*; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D010984:Platinum; D011137:Polystyrenes / Q000737:chemistry / Q000378:metabolism; D013053:Spectrophotometry / Q000379:methods*; D014025:Titanium
D011137:Polystyrenes; D010984:Platinum; D014025:Titanium