Bestman, J. E., Ewald, R. C., Chiu, S. L., Cline, H. T. (2006) In vivo single-cell electroporation for transfer of DNA and macromolecules. Nat Protoc, 1 (3). pp. 1267-72. ISSN 1750-2799 (Electronic)
Abstract
Single-cell electroporation allows transfection of plasmid DNA or macromolecules into individual living cells using modified patch electrodes and common electrophysiological equipment. This protocol is optimized for rapid in vivo electroporation of Xenopus laevis tadpole brains with DNA, dextrans, morpholinos and combinations thereof. Experienced users can electroporate roughly 40 tadpoles per hour. The technique can be adapted for use with other charged transfer materials and in other systems and tissues where cells can be targeted with a micropipette. Under visual guidance, an electrode filled with transfer material is placed in a cell body-rich area of the tadpole brain and a train of voltage pulses applied, which electroporates a nearby cell. We show examples of successfully electroporated single cells, instances of common problems and troubleshooting suggestions. Single-cell electroporation is an affordable method to fluorescently label and genetically manipulate individual cells. This powerful technique enables observation of single cells in an otherwise normal environment.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | Animals Brain metabolism DNA metabolism Electroporation methods Larva Plasmids metabolism Transfection methods Xenopus laevis |
| Subjects: | Investigative techniques and equipment > electrophysiology Investigative techniques and equipment > transfection > single cell electroporation Investigative techniques and equipment > transfection organism description > animal > Frog > xenopus |
| CSHL Authors: | |
| Communities: | CSHL labs > Cline lab School of Biological Sciences > Publications |
| Depositing User: | CSHL Librarian |
| Date: | 2006 |
| Date Deposited: | 20 Dec 2011 19:01 |
| Last Modified: | 19 Sep 2014 16:08 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/22753 |
Actions (login required)
 |
Administrator's edit/view item |