title: Nanofountain probe electroporation (NFP-E) of single cells.
abstract: The ability to precisely deliver molecules into single cells is of great interest to biotechnology researchers for advancing applications in therapeutics, diagnostics, and drug delivery toward the promise of personalized medicine. The use of bulk electroporation techniques for cell transfection has increased significantly in the past decade, but the technique is nonspecific and requires high voltage, resulting in variable efficiency and low cell viability. We have developed a new tool for electroporation using nanofountain probe (NFP) technology, which can deliver molecules into cells in a manner that is highly efficient and gentler to cells than bulk electroporation or microinjection. Here we demonstrate NFP electroporation (NFP-E) of single HeLa cells within a population by transfecting them with fluorescently labeled dextran and imaging the cells to evaluate the transfection efficiency and cell viability. Our theoretical analysis of the mechanism of NFP-E reveals that application of the voltage creates a localized electric field between the NFP cantilever tip and the region of the cell membrane in contact with the tip. Therefore, NFP-E can deliver molecules to a target cell with minimal effect of the electric potential on the cell. Our experiments on HeLa cells confirm that NFP-E offers single cell selectivity, high transfection efficiency (>95%), qualitative dosage control, and very high viability (92%) of transfected cells.
authors: Department of Mechanical Engineering, Northwestern University, Evanston, Illinois 60208, USA.
authors: Wonmo
authors: W
authors: Kang
authors: Fazel
authors: F
authors: Yavari
authors: Majid
authors: M
authors: Minary-Jolandan
authors: Juan P
authors: JP
authors: Giraldo-Vela
authors: Asmahan
authors: A
authors: Safi
authors: Rebecca L
authors: RL
authors: McNaughton
authors: Victor
authors: V
authors: Parpoil
authors: Horacio D
authors: HD
authors: Espinosa
mesh_terms: D018274:Electroporation; D036103:Nanotechnology; D059010:Single-Cell Analysis
references: Proc Natl Acad Sci U S A. 2008 Oct 28;105(43):16438-43
references: 18946047
references: Nat Protoc. 2009;4(6):862-9
references: 19444243
references: Cell. 2010 May 14;141(4):559-63
references: 20478246
references: Nat Rev Mol Cell Biol. 2011 Feb;12(2):119-25
references: 21224886
references: Biophys J. 2007 Jan 15;92(2):404-17
references: 17056739
references: Differentiation. 2002 Jun;70(4-5):148-54
references: 12147134
references: Biophys J. 2000 Oct;79(4):1993-2001
references: 11023903
references: Nat Methods. 2011 Apr;8(4):311-4
references: 21451520
references: Proc Natl Acad Sci U S A. 2010 Mar 30;107(13):5815-20
references: 20212151
references: Anal Chem. 2010 Jan 1;82(1):353-8
references: 19961232
references: Nat Methods. 2010 Apr;7(4):275-85
references: 20354518
references: PLoS One. 2011;6(7):e21687
references: 21799744
references: Bioessays. 2008 Jun;30(6):606-10
references: 18478541
references: Proc Natl Acad Sci U S A. 2007 May 15;104(20):8218-22
references: 17485677
references: Anal Chem. 2007 May 15;79(10):3589-96
references: 17444611
references: J Biomech. 2000 Jan;33(1):15-22
references: 10609514
references: Anal Chem. 2004 Dec 1;76(23):7045-52
references: 15571358
references: Nat Nanotechnol. 2011 Jan;6(1):57-64
references: 21151109
references: Pflugers Arch. 2002 Feb;443(4):664-70
references: 11907835
references: Langmuir. 2007 Aug 14;23(17):9120-3
references: 17645361
references: Curr Opin Biotechnol. 2004 Feb;15(1):44-9
references: 15102465
references: Science. 2011 Jan 7;331(6013):24-6
references: 21212334
references: Anal Chem. 2007 Dec 15;79(24):9584-7
references: 18004820
references: Nat Nanotechnol. 2012 Feb 12;7(3):185-90
references: 22327876
references: Proc Natl Acad Sci U S A. 2007 Jan 16;104(3):727-32
references: 17213326
references: Small. 2009 Jul;5(14):1667-74
references: 19437464
references: Anal Chem. 2001 Sep 15;73(18):4469-77
references: 11575795
references: Proc Natl Acad Sci U S A. 2001 Apr 10;98(8):4295-8
references: 11274366
references: Biochem Biophys Res Commun. 2005 Jul 8;332(3):633-9
references: 15925564
references: Lab Chip. 2008 Jan;8(1):62-7
references: 18094762
references: Anal Chem. 2007 Jun 15;79(12):4410-8
references: 17511419
references: Biochim Biophys Acta. 2005 Aug 5;1724(3):270-80
references: 15951114
references: FEBS Lett. 1974 Mar 1;39(3):249-51
references: 4368819
references: Proc Natl Acad Sci U S A. 1998 Sep 1;95(18):10356-60
references: 9724707
references: Lab Chip. 2005 Jan;5(1):38-43
references: 15616738
references: Nat Mater. 2011 May;10(5):389-97
references: 21499315
references: Proc Natl Acad Sci U S A. 2010 Feb 2;107(5):1870-5
references: 20080678
references: Nat Protoc. 2006;1(3):1267-72
references: 17406410
references: Neuron. 2001 Mar;29(3):583-91
references: 11301019
references: Nat Nanotechnol. 2011 Oct 16;6(11):747-54
references: 22002097
references: Nanomedicine. 2008 Sep;4(3):215-25
references: 18501680
references: Anal Bioanal Chem. 2010 Aug;397(8):3235-48
references: 20496058
references: Integr Biol (Camb). 2009 Mar;1(3):242-51
references: 20023735
references: Biosens Bioelectron. 2008 Apr 15;23(9):1353-60
references: 18242073
references: EMBO J. 1982;1(7):841-5
references: 6329708
references: Nano Lett. 2012 Aug 8;12(8):3881-6
references: 22166016
references: Lab Chip. 2004 Apr;4(2):104-8
references: 15052348
references: Lab Chip. 2005 Jan;5(1):23-9
references: 15616736
references: Nano Lett. 2009 May;9(5):2193-8
references: 19366190
references: Lab Chip. 2007 Apr;7(4):457-62
references: 17389961
references: Anal Bioanal Chem. 2006 Jun;385(3):474-85
references: 16534574