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Chapter 5
Tutorial
Guidelines for Single-Cell Electroporation
Successful single-cell electroporation is highly dependent on three factors: the
geometry of the glass micropipette tip, proximity between the micropipette tip and
the target cell, and the stimulation parameters.
Micropipettes
Fabricating Micropipettes
In general, tip diameter should be much smaller than the cell diameter, in the
range of 0.5 µm. The geometry of the micropipette tip shank may also be an
important consideration.
Micropipettes of the type described here will work for almost any cell type tried
to date. Use capillary glass whose internal diameter is 1.15 to 1.2 mm and
outside diameter is 1.6 to 1.7 mm with a glass filament fused to its interior to
facilitate tip filling. Inexpensive glass like Corning Pyrex or Kimble Kimax is
available from a variety of major suppliers. Purchase 4" pieces to pull two
similar length micropipettes of adequate length. It is recommended that the
back ends be flame-polished to prevent the sharp end from cutting the washer
within the micropipette holder.
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