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

Institute of Nano Engineering and Microsystems, National Tsing Hua University, Hsinchu City, Taiwan.
Biomedical Microdevices (Impact Factor: 2.72). 06/2012; 14(5):811-7. DOI: 10.1007/s10544-012-9660-9
Source: PubMed

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.

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    ABSTRACT: Despite the significant research in electroporation, high electric field was applied to the whole cells resulted in permeabilizing the membrane of millions of cells without reversibility [1]. To deliver biomolecules through the specific region of the cell membrane with high cell viability and high transfection rate is important for various biological and therapeutic applications. This report presents a new type localized single cell membrane electroporation (LSCMEP), at specific region of the single cell with the application of 800 µs electric pulse. The ITO nano-electrodes with 100nm thickness and 500 nm gap between two electrodes can generate an intense electric field to track biomolecules inside HeLa cell in our studies. This small gap between two nano-electrodes can neglect thermal effect on cell membrane and permit reversible electroporation with high cell viability (90%) and minimum effected electroporation region (0.48 µm). Our approach successfully delivers biomolecules through a specific region of single cell with high transfection rate (82%) and high cell viability. This process, not only generates well-controlled nano-pores allowing rapid recovery of cell membrane, but also it provides a clear optical path potentially tracking of drugs to deliver inside single cell.
    IEEE-NEMS-2013; 04/2013
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    ABSTRACT: The behaviors of cell to cell or cell to environment with their organelles and their intracellular physical or biochemical effects are still not fully understood. Analyzing millions of cells together cannot provide detailed information, such as cell proliferation, differentiation or different responses to external stimuli and intracellular reaction. Thus, single cell level research is becoming a pioneering research area that unveils the interaction details in high temporal and spatial resolution among cells. To analyze the cellular function, single cell electroporation can be conducted by employing a miniaturized device, whose dimension should be similar to that of a single cell. Micro/nanofluidic devices can fulfill this requirement for single cell electroporation. This device is not only useful for cell lysis, cell to cell fusion or separation, insertion of drug, DNA and antibodies inside single cell, but also it can control biochemical, electrical and mechanical parameters using electroporation technique. This device provides better performance such as high transfection efficiency, high cell viability, lower Joule heating effect, less sample contamination, lower toxicity during electroporation experiment when compared to bulk electroporation process. In addition, single organelles within a cell can be analyzed selectively by reducing the electrode size and gap at nanoscale level. This advanced technique can deliver (in/out) biomolecules precisely through a small membrane area (micro to nanoscale area) of the single cell, known as localized single cell membrane electroporation (LSCMEP). These articles emphasize the recent progress in micro/nanofluidic single cell electroporation, which is potentially beneficial for high-efficient therapeutic and delivery applications or understanding cell to cell interaction
    Journal of Micromachine. 09/2013; 4:333-356.


Available from
May 15, 2014