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

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Single-cell electroporation allows transfection of plasmid DNA or macrocmolecules 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.

*Note: In the version of this article initially published online, the article’s page numbers should have been 1267–1272. This error has been corrected in the PDF version of the article.

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Figure 1: Illustration of SCE in the optic tectum of the Xenopus laevis central nervous system.
Figure 2: The single-cell electroporation setup.
Figure 3: Electrode loading syringe.
Figure 4: Anticipated results of single-cell electroporation.
Figure 5: Examples of electroporation results requiring troubleshooting.

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  • 30 November 2006

    In the version of this article initially published online, the article’s page numbers should have been 1267–1272. This error has been corrected in the PDF version of the article.


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Correspondence to Hollis T Cline.

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Bestman, J., Ewald, R., Chiu, SL. et al. In vivo single-cell electroporation for transfer of DNA and macromolecules. Nat Protoc 1, 1267–1272 (2006).

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