Delivery of molecules into cells using localized single cell electroporation on ITO micro-electrode based transparent chip

Abstract

Single cell electroporation is one of the nonviral method which successfully allows transfection of exogenous macromolecules into individual living cell. We present localized cell membrane electroporation at single-cell level by using indium tin oxide (ITO) based transparent micro-electrodes chip with inverted microscope. A focused ion beam (FIB) technique has been successfully deployed to fabricate transparent ITO micro-electrodes with submicron gaps, which can generate more intense electric field to produce very localized cell membrane electroporation. In our approach, we have successfully achieved 0.93 μm or smaller electroporation region on the cell surface to inject PI (Propidium Iodide) dye into the cell with 60 % cell viability. This experiments successfully demonstrate the cell self-recover process from the injected PI dye intensity variation. Our localized cell membrane electroporation technique (LSCMEP) not only generates reversible electroporation process but also it provides a clear optical path for potentially monitoring/tracking of drugs to deliver in single cell level.

Tuning nano electric field to affect restrictive membrane area on localized single cell nano-electroporation

“…Recently single cell membrane nano-electroporation in a localized region has attracted much attention due to high transfection rate and high cell viability. [8][9][10][11][12] However the effect of highly intensive electric field with the variation of TMP in a local region of the single cell membrane was not well studied previously. Higher electric field effects the whole single cell (familiar as single cell electroporation (SCEP)) to cause cell lysis easily.…”

Recent Trends on Micro/Nanofluidic Single Cell Electroporation

“…To apply a very high electric field in between two microchannels, a transfection agent was delivered through the nanochannel using an electrophoretically driven process and finally drugs were delivered inside a single cell through a very small area of the cell membrane. In 2012, Chen et al demonstrated another localized single cell membrane electroporation usimg ITO microelectrode based transparent chip [27]. Figure 14 shows microfluidic localized single cell membrane electroporation device.…”

“…As a result, single cell manipulation can be performed from population of cells together. These devices can analyze cell to cell behavior with their organelle, their orientation, changes of cell size and shape with different polarities of electric fields [14,23,24,26,27]. On the other hand, bulk electroporation needs two large electrodes surrounding millions of cells together and a homogeneous electric field is applied to electroporates millions of cells at once.…”

“…To use this dimensional advantage, an electric field can easily intense in the local region of the single cell membrane, by which high transfection rate and high cell viability were achieved [27,29]. However localized electroporation can also be performed by nanochannel ion transportation using the electrophoresis method [30].…”

“…However localized electroporation can also be performed by nanochannel ion transportation using the electrophoresis method [30]. To apply electrical field in local region of the single cell membrane, the process is known as localized single cell membrane electroporation (LSCMEP), by which precise and controllable drug delivery is possible [27,29,30]. The LSCMEP process can provide very high transfection rate, high cell viability, low power consumption, lower toxicity, low thermal effect in comparison with SCEP or BEP process.…”