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.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Bestman JE,Ewald RC,Chiu SL,Cline HTdoi
10.1038/nprot.2006.186subject
Has Abstractpub_date
2006-01-01 00:00:00pages
1267-72issue
3eissn
1754-2189issn
1750-2799pii
nprot.2006.186journal_volume
1pub_type
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