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The authors describe an efficient technique for the selective chemical and biological …


Biology Articles » Biophysics » Altering the biochemical state of individual cultured cells and organelles with ultramicroelectrodes

Abstract
- Altering the biochemical state of individual cultured cells and organelles with ultramicroelectrodes

Altering the biochemical state of individual cultured cells and organelles with ultramicroelectrodes
 
J. Anders Lundqvist*, Frida Sahlin*,dagger, Maria A. I. Åbergdagger,Dagger,Anette Strömberg*, Peter S. ErikssonDagger, and Owe Orwar*

* Department of Chemistry, Göteborg University, SE-412 96 Göteborg, Sweden; and Dagger Department of Neurology, Institute of Clinical Neuroscience, Sahlgrenska University Hospital, SE-413 45 Göteborg, Sweden

Communicated by Richard N. Zare, Stanford University, Stanford, CA, July 6, 1998 (received for review February 20, 1998)

We describe an efficient technique for the selective chemical and biological manipulation of the contents of individual cells. This technique is based on the electric-field-induced permeabilization (electroporation) in biological membranes using a low-voltage pulse generator and microelectrodes. A spatially highly focused electric field allows introduction of polar cell-impermeant solutes such as fluorescent dyes, fluorogenic reagents, and DNA into single cells. The high spatial resolution of the technique allows for design of, for example, cellular network constructions in which cells in close contact with each other can be made to possess different biochemical, biophysical, and morphological properties. Fluorescein, and fluo-3 (a calcium-sensitive fluorophore), are electroporated into the soma of cultured single progenitor cells derived from adult rat hippocampus. Fluo-3 also is introduced into individual submicrometer diameter processes of thapsigargin-treated progenitor cells, and a plasmid vector cDNA construct (pRAY 1), expressing the green fluorescent protein, is electroporated into cultured single COS 7 cells. At high electric field strengths, observations of dye-transfer into organelles are proposed.

Source: Proc Natl Acad Sci U S A.  Vol. 95, Issue 18, 10356-10360, September 1, 1998.


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