The design and fabrication of a localized single cell electroporation chip
| Autor: | Chen, Sheng-Chieh, 陳聖杰 |
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| Rok vydání: | 2010 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 98 Tracking a single molecule in the cell plays a significant role in biology. Currently, there is a need for a method to deliver molecules to a cell while tracking such delivery. We develop a new platform for tracking a single molecule delivered by localized eletropoation. In this work, a single cell is elecroporated locally at a specific membrane position on a transparent electrode chip. The whole eletroporation process was recorded by CCD camera by tracking PI dye entering the HeLa cell. Transparent ITO electrodes were employed to apply a confine E-field in a sub-micron range for localized cell electroporation. In conventional single cell electroporation, electric field are applied to the whole cell between tow electrode. As a result many pores will be generated with different permeability and size, resulting in difficulties in tracking molecules delivered into the cell. In this paper, very localized electroporation at a specified region on a single cell surface by transparent ITO electrodes is realized, which can not only generate well-controlled nano-pores allowing for rapid recovery of the cell surface, but also provide a clear optical path for cell monitoring and molecular tracking. In this work we demonstrate very localized electroporation to decrease the damage to the cell membrane. We can also observe resealing of the electroporation hole. We use a diffusion model to confirm localized electroportion is confined in 0.9μm region. We can control electroporation region down to 0.9μm and control the electroporation time to increase the survivability. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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