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Biophys J. 1998 Nov;75(5):2558-63.

Lipid-assisted microinjection: introducing material into the cytosol and membranes of small cells.

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1
Molecular Signaling Group, University Department of Surgery, University of Wales College of Medicine, Heath Park, Cardiff CF4 4XN, Wales.

Abstract

The microinjection of synthetic molecules, proteins, and nucleic acids into the cytosol of living cells is a powerful technique in cell biology. However, the insertion of a glass micropipette into the cell is a potentially damaging event, which presents significant problems, especially for small mammalian cells (spherical diameter = 2-15 micron), especially if they are only loosely adherent. The current technique is therefore limited to cells that are both sufficiently large or robust and firmly attached to a substrate. We describe here a modification of the standard technique that overcomes some of the problems associated with conventional microinjection but that does not involve the insertion of a micropipette deep into the cell cytoplasm. Instead, this method depends on lipid fusion at the micropipette tip to form a continuous but temporary conductance pathway between the interiors of the micropipette and cell. This technique thus also provides a novel method of transferring lipids and lipid-associated molecules to the plasma membrane of cells.

PMID:
9788951
PMCID:
PMC1299930
DOI:
10.1016/S0006-3495(98)77700-8
[Indexed for MEDLINE]
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