Abstract
Electroendocytosis (EED), i.e. electric field-induced endocytosis, is a technique for bio-molecule and drug delivery to cells using a pulsed electric field lower than that applied in electroporation (EP). Different from EP in which nanometer-sized electropores appear on the plasma membrane lipid bilayer, EED induces cell membrane internalization and fission via endocytotic vesicles. In this study, we conduct comprehensive experimental study on the EED of HeLa cells using a micro chip and the corresponding endocytotic vesicles were visualized and investigated by using FM4-64 fluorescent dye and in situ fluorescence microscopy. The uptake of molecules by the EED of cells was characterized by average intracellular fluorescent intensity from a large number (>2,000) of single cells. The EED efficiency was determined as a function of three electric parameters (electric field strength, pulse duration, total electric treatment time). The EED efficiency as a function of electric field strength clearly shows biphasic characteristics at different experimental conditions. The EED experiments using cytoskeleton inhibitors illustrate unique mechanisms distinct from EP. This study provides a foundation for further on-chip study of the time-dependent mechanism of EED at the single-cell level.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
H. Andersson, A. van den Berg, Sensor Actuat B-Chem 92(3), 315–325 (2003)
H. Andersson, A. van den Berg, Curr Opin Biotechnol 15(1), 44–49 (2004)
Y. Antov, A. Barbul, R. Korenstein, Exp Cell Res 297(2), 348–362 (2004)
Y. Antov, A. Barbul, H. Mantsur, R. Korenstein, Biophys J 88(3), 2206–2223 (2005)
G. Apodaca, Traffic 2(3), 149–159 (2001)
D.C. Bartoletti, G.I. Harrison, J.C. Weaver, FEBS Lett 256(1–2), 4–10 (1989)
S. Bolte, C. Talbot, Y. Boutte, O. Catrice, N.D. Read, B. Satiat-Jeunemaitre, J Microsc-Oxford 214(2), 159–173 (2004)
P.J. Canatella, J.F. Karr, J.A. Petros, M.R. Prausnitz, Biophys J 80(2), 755–764 (2001)
D.C. Chang, B.M. Chassy, J.A. Saunders, A.E. Sowers, Guide to electroporation and electrofusion (Academic, San Diego, 1992), pp. 9–27
S.D. Conner, S.L. Schmid, Nature 422(6927), 37–44 (2003)
J.D. Deng, K.H. Schoenbach, E.S. Buescher, P.S. Hair, P.M. Fox, S.J. Beebe, Biophys J 84(4), 2709–2714 (2003)
M.B. Fox, D.C. Esveld, A. Valero, R. Luttge, H.C. Mastwijk, P.V. Bartels, A. van den Berg, R.M. Boom, Anal Bioanal Chem 385(3), 474–485 (2006)
M.A. Gaffield, W.J. Betz, Nat Protoc 1(6), 2916–2921 (2006)
M. Glogauer, W. Lee, C.A.G. Mcculloch, Exp Cell Res 208(1), 232–240 (1993)
M. Golzio, J. Teissie, M.P. Rols, Proc Natl Acad Sci 99(3), 1292–1297 (2002)
R. P. Haugland, Handbook of fluorescent probes and research products, (Molecular Probes, Eugene, 2002), pp. 578, 683–684
H.Q. He, D.C. Chang, Y.K. Lee, Bioelectrochemistry 68(1), 89–97 (2006)
H.Q. He, D.C. Chang, Y.K. Lee, Bioelectrochemistry 70(2), 363–368 (2007)
H.Q. He, D.C. Chang, Y.K. Lee, Bioelectrochemistry 72(2), 161–168 (2008)
Y. Huang, B. Rubinsky, Sensor Actuat a-Phys 89(3), 242–249 (2001)
M. Jin, M.D. Snider, J Biol Chem 268(24), 18390–18397 (1993)
M. Khine, A. Lau, C. Ionescu-Zanetti, J. Seo, L.P. Lee, Lab Chip 5(1), 38–43 (2005)
W. Krassowska, P.D. Filev, Biophys J 92(2), 404–417 (2007)
S. Kumari, S. Mg, S. Mayor, Cell Res 20(3), 256–275 (2010)
H. Lambert, R. Pankov, J. Gauthier, R. Hancock, Biochem Cell Biol 68(4), 729–734 (1990)
I. Mellman, Annu Rev Cell Dev Biol 12(1), 575–625 (1996)
E. Neumann, A.E. Sowers, C.A. Jordan, Electroporation and electrofusion in cell biology (Plenum Press, New York, 1989), pp. 61–82
W.D. Niles, A.B. Malik, J Membr Biol 167(1), 85–101 (1999)
H. Noguchi, M. Matsushita, S. Matsumoto, Y.F. Lu, H. Matsui, S. Bonner-Weir, Biochem Biophys Res Commun 332(1), 68–74 (2005)
C.Y. Okada, M. Rechsteiner, Cell 29(1), 33–41 (1982)
A. Piasek, J. Thyberg, J Cell Sci 45(1), 59–71 (1980)
Z.M. Qian, H.Y. Li, H.Z. Sun, K. Ho, Pharmacol Rev 54(4), 561–587 (2002)
M.P. Rols, P. Femenia, J. Teissie, Biochem Biophys Res Commun 208(1), 26–35 (1995)
Y. Rosemberg, R. Korenstein, Bioelectrochem Bioenerg 42(2), 275–281 (1997)
T.A. Vida, S.D. Emr, J Cell Biol 128(5), 779–792 (1995)
M.E. Ward, A. Murray, J Gen Microbiol 130(7), 1765–1780 (1984)
J.N. Weinstein, S. Yoshikami, P. Henkart, R. Blumenthal, W.A. Hagins, Science 195(4277), 489–492 (1977)
G.M. Whitesides, Nature 442(7101), 368–373 (2006)
C.J.G. Yeh, B.L. Hsi, W.P. Faulk, J Immunol Methods 43(3), 269 (1981)
U. Zimmermann, R. Schnettler, G. Klock, H. Watzka, E. Donath, R.W. Glaser, Naturwissenschaften 77(11), 543–545 (1990)
Acknowledgements
This work was supported by the Hong Kong Research Grants Council (Project Ref No. 615907). The authors would like to thank Mr. Guangyao Yin, Mr. Wentao Wang, Dr. Peigang Deng, Ms. Inez Tsui, Mr. Wan Lap Yeung, and Mr. Allen Ng at HKUST for technical support.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Lin, R., Chang, D.C. & Lee, YK. Single-cell electroendocytosis on a micro chip using in situ fluorescence microscopy. Biomed Microdevices 13, 1063–1073 (2011). https://doi.org/10.1007/s10544-011-9576-9
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10544-011-9576-9