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1.
Bioelectrochemistry ; 102: 35-41, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25483998

RESUMO

Electroporation is a commonly used approach to rapidly introduce exogenous molecules into cells without permanent damage. Compared to classical electroporation protocols, microchip-based electroporation approaches have the advantages of high transfection efficiency and low consumption, but they also commonly rely on costly and tedious microfabrication technology. Hence, it is desirable to develop a novel, more affordable, and effective approach to facilitate cell electroporation. In this study, we utilized a standard printed circuit board (PCB) technology to fabricate a chip with an interdigitated array of electrodes for electroporation of suspended cells. The electrodes (thickness ~35 µm) fabricated by PCB technology are much thicker than the two-dimensional (2D) planar electrodes (thickness < 1 µm) fabricated by conventional microfabrication techniques and possess a smooth corner edge. Numerical simulations showed that the three-dimensional (3D) electrodes fabricated by PCB technology can provide a more uniformly distributed electric field compared to 2D planar electrodes, which is beneficial for reducing the electrolysis of water and improving cell transfection efficiency. The chip constructed here is composed of 18 individually addressable wells for high throughput cell electroporation. HeLa, MCF7, COS7, Jurkat, and 3T3-L1 cells were efficiently transfected with the pEGFP-N1 plasmid using individually optimal electroporation parameters. This work provides a novel method for convenient and rapid cell transfection and thus holds promise for use as a low-cost disposable device in biomedical research.


Assuntos
Eletroporação/instrumentação , Animais , Linhagem Celular Tumoral , Eletroporação/economia , Proteínas de Fluorescência Verde/genética , Humanos , Camundongos , Microeletrodos , Plasmídeos/genética , Transfecção
2.
Braz. j. med. biol. res ; 46(9): 752-757, 19/set. 2013. graf
Artigo em Inglês | LILACS | ID: lil-686571

RESUMO

One of the challenges of the postgenomic era is characterizing the function and regulation of specific genes. For various reasons, the early chick embryo can easily be adopted as an in vivo assay of gene function and regulation. The embryos are robust, accessible, easily manipulated, and maintained in the laboratory. Genomic resources centered on vertebrate organisms increase daily. As a consequence of optimization of gene transfer protocols by electroporation, the chick embryo will probably become increasingly popular for reverse genetic analysis. The challenge of establishing chick embryonic electroporation might seem insurmountable to those who are unfamiliar with experimental embryological methods. To minimize the cost, time, and effort required to establish a chick electroporation assay method, we describe and illustrate in great detail the procedures involved in building a low-cost electroporation setup and the basic steps of electroporation.


Assuntos
Animais , Embrião de Galinha , Eletroporação/economia , Eletroporação/instrumentação , Eletroporação/métodos , Regulação da Expressão Gênica/genética , Técnicas de Transferência de Genes/instrumentação , Eletrodos , Desenho de Equipamento , Proteínas de Fluorescência Verde
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