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.
Full text
PDF
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bakker Schut T. C., Florians A., Radosević K., de Grooth B. G., Greve J. Signal processing in slit-scan flow cytometry of cell conjugates. Cytometry. 1993;14(4):459–464. doi: 10.1002/cyto.990140417. [DOI] [PubMed] [Google Scholar]
- Bakker Schut T. C., de Grooth B. G., Greve J. A new principle of cell sorting by using selective electroporation in a modified flow cytometer. Cytometry. 1990;11(6):659–666. doi: 10.1002/cyto.990110602. [DOI] [PubMed] [Google Scholar]
- Lo M. M., Tsong T. Y., Conrad M. K., Strittmatter S. M., Hester L. D., Snyder S. H. Monoclonal antibody production by receptor-mediated electrically induced cell fusion. 1984 Aug 30-Sep 5Nature. 310(5980):792–794. doi: 10.1038/310792a0. [DOI] [PubMed] [Google Scholar]
- Tomita M., Tsong T. Y. Selective production of hybridoma cells: antigenic-based pre-selection of B lymphocytes for electrofusion with myeloma cells. Biochim Biophys Acta. 1990 Dec 10;1055(3):199–206. doi: 10.1016/0167-4889(90)90033-a. [DOI] [PubMed] [Google Scholar]
- Zimmermann U. Electric field-mediated fusion and related electrical phenomena. Biochim Biophys Acta. 1982 Nov 30;694(3):227–277. doi: 10.1016/0304-4157(82)90007-7. [DOI] [PubMed] [Google Scholar]