In vivo single-cell electroporation for transfer of DNA and macromolecules

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)

URL: http://www.ncbi.nlm.nih.gov/pubmed/17406410
DOI: 10.1038/nprot.2006.186

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
Watson School > Publications
Depositing User: CSHL Librarian
Date: 2006
Date Deposited: 20 Dec 2011 19:01
Last Modified: 19 Sep 2014 16:08
Related URLs:
URI: http://repository.cshl.edu/id/eprint/22753

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