Selective Electrofusion of Conjugated Cells in Flow

Tom C. Bakker Schut, Yvonne M. Kraan, Wim Barlag, Lou de Leij, Bart G. de Grooth, Jan Greve

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    Abstract

    Using a modified flow cytometer we have induced electrofusion of K562 and L1210 cells in flow. The two cell types are stained with two different fluorescent membrane probes, DiO and DiI, to facilitate optical recognition, and then coupled through an avidin-biotin bridge. In the flow cytometer, the hydrodynamically focused cells and cell pairs are first optically analyzed in a normal flow channel and then forced to flow through a Coulter orifice. If the optical analysis indicates that a cell pair is present, an electric pulse is applied across the orifice to induce fusion. The pulsed cell pairs were subsequently analyzed using normal and confocal microscopy to evaluate fusion induction. It appears that fusion can be induced in about 10% of pulsed cell pairs when one electric pulse with a duration of 10–15 microseconds and an effective electric field strength of 4–8 10(5) V/m is used.
    Original languageEnglish
    Pages (from-to)568-572
    Number of pages5
    JournalBiophysical journal
    Volume65
    Issue number2
    DOIs
    Publication statusPublished - 1993

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