title: Survival rate of eukaryotic cells following electrophoretic nanoinjection.
abstract: Insertion of foreign molecules such as functionalized fluorescent probes, antibodies, or plasmid DNA to living cells requires overcoming the plasma membrane barrier without harming the cell during the staining process. Many techniques such as electroporation, lipofection or microinjection have been developed to overcome the cellular plasma membrane, but they all result in reduced cell viability. A novel approach is the injection of cells with a nanopipette and using electrophoretic forces for the delivery of molecules. The tip size of these pipettes is approximately ten times smaller than typical microinjection pipettes and rather than pressure pulses as delivery method, moderate DC electric fields are used to drive charged molecules out of the tip. Here, we show that this approach leads to a significantly higher survival rate of nanoinjected cells and that injection with nanopipettes has a significantly lower impact on the proliferation behavior of injected cells. Thus, we propose that injection with nanopipettes using electrophoretic delivery is an excellent alternative when working with valuable and rare living cells, such as primary cells or stem cells.
authors.affiliation: Biomolecular Photonics, Department of Physics, University of Bielefeld, Universitätsstr. 25, 33615 Bielefeld Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Simonis
authors.affiliation: Biomolecular Photonics, Department of Physics, University of Bielefeld, Universitätsstr. 25, 33615 Bielefeld Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Hübner
authors.affiliation: Biomolecular Photonics, Department of Physics, University of Bielefeld, Universitätsstr. 25, 33615 Bielefeld Germany.
authors.forename: Alice
authors.initials: A
authors.lastname: Wilking
authors.affiliation: Biomolecular Photonics, Department of Physics, University of Bielefeld, Universitätsstr. 25, 33615 Bielefeld Germany.|Department of Internal Medicine and NSF Center for Biophotonics, University of California, Davis, Sacramento, CA 95817, USA.
authors.forename: Thomas
authors.initials: T
authors.lastname: Huser
authors.affiliation: Biomolecular Photonics, Department of Physics, University of Bielefeld, Universitätsstr. 25, 33615 Bielefeld Germany.
authors.forename: Simon
authors.initials: S
authors.lastname: Hennig
mesh_terms: D002232:Carbocyanines / Q000378:metabolism; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival; D004560:Electricity; D004586:Electrophoresis; D005057:Eukaryotic Cells / Q000166:cytology* / Q000378:metabolism; D005453:Fluorescence; D006801:Humans; D008845:Microinjections / Q000379:methods*; D036103:Nanotechnology / Q000379:methods*; D013997:Time Factors; D059008:Time-Lapse Imaging
chemical_list: C569686:Alexa Fluor 647; D002232:Carbocyanines