Real-time monitoring of cell viability using direct electrical measurement with a patch-clamp microchip

Abstract

Real-time tagless monitoring of cell viability using patch-clamp microchips is reported and validated by using fluorescence imaging techniques for the first time. Specifically, four human breast cancer cell lines (MDA-MB231, MDA-MB231-brain metastatic subline (abbreviated as MB231-BR), MB231-BR over-expressing HER2 gene (MB231-BR-HER2), and MB231-BR-vector control for the HER2 (MB231-BR-vector)) have been used for these studies. Systematic experiments on these cells found that the seal impedance/resistance of cells captured by the micro-pipettes always decreases during the process when the cell loses its viability, and therefore it is a valid indicator of live or dead cells. Systematic experiments also found that the Mega-seal of patch-clamp microchip is sufficient for monitoring cell viability. Given its simplicity of direct electrical measurement of cells without fluorescence labeling, this technology may provide an efficient technical platform to monitor the drug effects on cells, thereby significantly benefiting high throughput drug screening and discovery process.

On-Chip Studies of Magnetic Stimulation Effect on Single Neural Cell Viability and Proliferation on Glass and Nanoporous Surfaces

“…Viability of N27 Cells on Different Substrates under MS. To assess the viability of N27 cells under MS, cellular viability tests by calcein AM and propidium iodide (PI)-based live/dead assay have been performed. 49 In these experiments, four substrates have been used, including coverslip glass, AAO substrate, glass microchip, and AAO microchip. All the surfaces of these substrates are coated with poly-D-lysine.…”

Simultaneous <i>in situ</i> Detection of the Optical Fluorescence, Fluorescence Recovery Kinetics After Photobleaching & Membrane Ion Flux on the Electrophysiological Lab-on-a-Chip

“…Thus, we designed a complex measurement system which, unlike the previously known systems for real-time patch-clamp on-a-chip, allows not only registration and screening [22] or a simultaneous optical detection and electrophysiological measurements [23], but also a comparison with the SIMULINK model with both the measurement and simulation data transmission to either a host machine or to the local network through the Ethernet protocol.…”

Prototype for Automatable, Dielectrophoretically-Accessed Intracellular Membrane–Potential Measurements by Metal Electrodes

“…Similarly to other devices based on the planar patch-clamp technique, the seal resistance measurement (also by impedance spectroscopy) could offer a standard output to gauge cell health. 41 In an automated system, however, the insulating matrix should exhibit good sealing properties. As known from the patch-clamp techniques, it would be desirable that the polymer behaves similarly to glass, so that a high seal resistance between cell membrane and polymer is facilitated, as it is known from Orcomer Ò or polydimethylsiloxane.…”