Abstract

The introduction and subsequent expression of foreign DNA inside living mammalian cells (transfection) is achieved by photoporation with a violet diode laser. We direct a compact 405 nm laser diode source into an inverted optical microscope configuration and expose cells to 0.3 mW for 40 ms. The localized optical power density of ~1200 MW/m2 is six orders of magnitude lower than that used in femtosecond photoporation (~104 TW/m2). The beam perforates the cell plasma membrane to allow uptake of plasmid DNA containing an antibiotic resistant gene as well as the green fluorescent protein (GFP) gene. Successfully transfected cells then expand into clonal groups which are used to create stable cell lines. The use of the violet diode laser offers a new and simple poration technique compatible with standard microscopes and is the simplest method of laser-assisted cell poration reported to date.

© 2005 Optical Society of America

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  1. F.L. Graham and A.J. Van der Eb, “A new technique for the assay of infectivity of human adenovirus 5 DNA,” Virology 52, 456–467 (1973).
    [CrossRef] [PubMed]
  2. R. Fraley, S. Subramani, P. Berg, and D. Papahadjopolous, “Introduction of liposome-encapsulated SV40 DNA into cells,” J. Biol. Chem. 255, 10431–10435 (1980).
    [PubMed]
  3. D.A. Rubinson, C.P. Dillon, A.V. Kwiatkowski, C. Sievers, L. Yang, J. Kopinja, M. Zhang, M.T. McManus, F.B. Gertler, M.L. Scott, and L. Van Parijs, “A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference,” Nature Genet. 33, 401–406 (2003).
    [CrossRef] [PubMed]
  4. T.K. Wong and E. Neumann, “Electric-field mediated gene-transfer,” Biochem. and Biophys. Res. Commun. 107, 584–587 (1982).
    [CrossRef]
  5. H. Schneckenburger, A. Hendinger, R. Sailer, W.S.L. Strauss, and M. Schmitt, “Laser-assisted optoporation of single cells,” J. Biomed. Opt. 7, 410–416 (2002).
    [CrossRef] [PubMed]
  6. G. Palumbo, M. Caruso, E. Crescenzi, M.F. Tecce, G. Roberti, and A. Colasanti, “Targeted gene transfer in eucaryotic cells by dye-assisted laser optoporation,” J. Photochem. Photobiol. B-Biol. 36, 41–46 (1996).
    [CrossRef]
  7. Y. Shirahata, N. Ohkohchi, H. Itagak, and S. Satomi, “New technique for gene transfection using laser irradiation,” J. Invest. Med. 49, 184–190 (2001).
    [CrossRef]
  8. J.S. Soughayer, T. Krasieva, S.C. Jacobson, J.M. Ramsey, B.J. Tromberg, and N.L. Allbritton, “Characterization of cellular optoporation with distance,” Anal. Chem. 72, 1342–1347 (2000).
    [CrossRef] [PubMed]
  9. S.K. Mohanty, M. Sharma, and P.K. Gupta, “Laser-assisted microinjection into targeted animal cells,” Biotechnol. Lett. 25, 895–899 (2003).
    [CrossRef] [PubMed]
  10. U.K. Tirlapur and K. Konig, “Femtosecond near-infrared laser pulses as a versatile non- invasive tool for intra-tissue nanoprocessing in plants without compromising viability,” Plant J. 31, 365–374 (2002).
    [CrossRef] [PubMed]
  11. U.K. Tirlapur and K. Konig, “Targeted transfection by femtosecond laser,” Nature 418, 290–291 (2002).
    [CrossRef] [PubMed]
  12. E. Zeira, A. Manevitch, A. Khatchatouriants, O. Pappo, E. Hyam, M. Darash-Yahana, E. Tavor, A. Honigman, A. Lewis, and E. Galun, “Femtosecond Infrared Laser - An Efficient and Safe in Vivo Gene Delivery System for Prolonged Expression,” Mol. Therapy 8, 342–350 (2003).
    [CrossRef]
  13. J.M. Girkin and A.I. Ferguson, “Confocal microscopy using an InGaN violet laser diode at 406nm,” Opt. Express 7, 336–341 (2000). http://www.opticsexpress.org/abstract.cfm?URI=OPEX-7-10-336
    [CrossRef] [PubMed]
  14. T.K. Lake, A.E. Carruthers, L. Paterson, M. Taylor, F. Gunn-Moore, J.W. Allen, W. Sibbett, and K. Dholakia, “Optical trapping and fluorescence excitation with violet diode lasers and extended cavity surface emitting lasers,” Opt. Express 12, 670–678 (2004). http://www.opticsexpress.org/abstract.cfm?URI=OPEX-12-4-670
    [CrossRef] [PubMed]
  15. P. Southern and P. Berg, “Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter,” J. Mol. Appl. Genet. 1, 327–341 (1982).
    [PubMed]
  16. Y Liu, D. K. Cheng, G. J. Sonek, M. W. Berns, C. F. Chapman, and B J. Tromberg, “Evidence for localized cell heating induced by infrared optical tweezers,” Biophys. J. 68, 2137–2144 (1995).
    [CrossRef] [PubMed]
  17. D. McGloin and K. Dholakia, private communication

2004 (1)

2003 (3)

S.K. Mohanty, M. Sharma, and P.K. Gupta, “Laser-assisted microinjection into targeted animal cells,” Biotechnol. Lett. 25, 895–899 (2003).
[CrossRef] [PubMed]

E. Zeira, A. Manevitch, A. Khatchatouriants, O. Pappo, E. Hyam, M. Darash-Yahana, E. Tavor, A. Honigman, A. Lewis, and E. Galun, “Femtosecond Infrared Laser - An Efficient and Safe in Vivo Gene Delivery System for Prolonged Expression,” Mol. Therapy 8, 342–350 (2003).
[CrossRef]

D.A. Rubinson, C.P. Dillon, A.V. Kwiatkowski, C. Sievers, L. Yang, J. Kopinja, M. Zhang, M.T. McManus, F.B. Gertler, M.L. Scott, and L. Van Parijs, “A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference,” Nature Genet. 33, 401–406 (2003).
[CrossRef] [PubMed]

2002 (3)

H. Schneckenburger, A. Hendinger, R. Sailer, W.S.L. Strauss, and M. Schmitt, “Laser-assisted optoporation of single cells,” J. Biomed. Opt. 7, 410–416 (2002).
[CrossRef] [PubMed]

U.K. Tirlapur and K. Konig, “Femtosecond near-infrared laser pulses as a versatile non- invasive tool for intra-tissue nanoprocessing in plants without compromising viability,” Plant J. 31, 365–374 (2002).
[CrossRef] [PubMed]

U.K. Tirlapur and K. Konig, “Targeted transfection by femtosecond laser,” Nature 418, 290–291 (2002).
[CrossRef] [PubMed]

2001 (1)

Y. Shirahata, N. Ohkohchi, H. Itagak, and S. Satomi, “New technique for gene transfection using laser irradiation,” J. Invest. Med. 49, 184–190 (2001).
[CrossRef]

2000 (2)

J.S. Soughayer, T. Krasieva, S.C. Jacobson, J.M. Ramsey, B.J. Tromberg, and N.L. Allbritton, “Characterization of cellular optoporation with distance,” Anal. Chem. 72, 1342–1347 (2000).
[CrossRef] [PubMed]

J.M. Girkin and A.I. Ferguson, “Confocal microscopy using an InGaN violet laser diode at 406nm,” Opt. Express 7, 336–341 (2000). http://www.opticsexpress.org/abstract.cfm?URI=OPEX-7-10-336
[CrossRef] [PubMed]

1996 (1)

G. Palumbo, M. Caruso, E. Crescenzi, M.F. Tecce, G. Roberti, and A. Colasanti, “Targeted gene transfer in eucaryotic cells by dye-assisted laser optoporation,” J. Photochem. Photobiol. B-Biol. 36, 41–46 (1996).
[CrossRef]

1995 (1)

Y Liu, D. K. Cheng, G. J. Sonek, M. W. Berns, C. F. Chapman, and B J. Tromberg, “Evidence for localized cell heating induced by infrared optical tweezers,” Biophys. J. 68, 2137–2144 (1995).
[CrossRef] [PubMed]

1982 (2)

P. Southern and P. Berg, “Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter,” J. Mol. Appl. Genet. 1, 327–341 (1982).
[PubMed]

T.K. Wong and E. Neumann, “Electric-field mediated gene-transfer,” Biochem. and Biophys. Res. Commun. 107, 584–587 (1982).
[CrossRef]

1980 (1)

R. Fraley, S. Subramani, P. Berg, and D. Papahadjopolous, “Introduction of liposome-encapsulated SV40 DNA into cells,” J. Biol. Chem. 255, 10431–10435 (1980).
[PubMed]

1973 (1)

F.L. Graham and A.J. Van der Eb, “A new technique for the assay of infectivity of human adenovirus 5 DNA,” Virology 52, 456–467 (1973).
[CrossRef] [PubMed]

Allbritton, N.L.

J.S. Soughayer, T. Krasieva, S.C. Jacobson, J.M. Ramsey, B.J. Tromberg, and N.L. Allbritton, “Characterization of cellular optoporation with distance,” Anal. Chem. 72, 1342–1347 (2000).
[CrossRef] [PubMed]

Allen, J.W.

Berg, P.

P. Southern and P. Berg, “Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter,” J. Mol. Appl. Genet. 1, 327–341 (1982).
[PubMed]

R. Fraley, S. Subramani, P. Berg, and D. Papahadjopolous, “Introduction of liposome-encapsulated SV40 DNA into cells,” J. Biol. Chem. 255, 10431–10435 (1980).
[PubMed]

Berns, M. W.

Y Liu, D. K. Cheng, G. J. Sonek, M. W. Berns, C. F. Chapman, and B J. Tromberg, “Evidence for localized cell heating induced by infrared optical tweezers,” Biophys. J. 68, 2137–2144 (1995).
[CrossRef] [PubMed]

Carruthers, A.E.

Caruso, M.

G. Palumbo, M. Caruso, E. Crescenzi, M.F. Tecce, G. Roberti, and A. Colasanti, “Targeted gene transfer in eucaryotic cells by dye-assisted laser optoporation,” J. Photochem. Photobiol. B-Biol. 36, 41–46 (1996).
[CrossRef]

Chapman, C. F.

Y Liu, D. K. Cheng, G. J. Sonek, M. W. Berns, C. F. Chapman, and B J. Tromberg, “Evidence for localized cell heating induced by infrared optical tweezers,” Biophys. J. 68, 2137–2144 (1995).
[CrossRef] [PubMed]

Cheng, D. K.

Y Liu, D. K. Cheng, G. J. Sonek, M. W. Berns, C. F. Chapman, and B J. Tromberg, “Evidence for localized cell heating induced by infrared optical tweezers,” Biophys. J. 68, 2137–2144 (1995).
[CrossRef] [PubMed]

Colasanti, A.

G. Palumbo, M. Caruso, E. Crescenzi, M.F. Tecce, G. Roberti, and A. Colasanti, “Targeted gene transfer in eucaryotic cells by dye-assisted laser optoporation,” J. Photochem. Photobiol. B-Biol. 36, 41–46 (1996).
[CrossRef]

Crescenzi, E.

G. Palumbo, M. Caruso, E. Crescenzi, M.F. Tecce, G. Roberti, and A. Colasanti, “Targeted gene transfer in eucaryotic cells by dye-assisted laser optoporation,” J. Photochem. Photobiol. B-Biol. 36, 41–46 (1996).
[CrossRef]

Darash-Yahana, M.

E. Zeira, A. Manevitch, A. Khatchatouriants, O. Pappo, E. Hyam, M. Darash-Yahana, E. Tavor, A. Honigman, A. Lewis, and E. Galun, “Femtosecond Infrared Laser - An Efficient and Safe in Vivo Gene Delivery System for Prolonged Expression,” Mol. Therapy 8, 342–350 (2003).
[CrossRef]

Dholakia, K.

Dillon, C.P.

D.A. Rubinson, C.P. Dillon, A.V. Kwiatkowski, C. Sievers, L. Yang, J. Kopinja, M. Zhang, M.T. McManus, F.B. Gertler, M.L. Scott, and L. Van Parijs, “A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference,” Nature Genet. 33, 401–406 (2003).
[CrossRef] [PubMed]

Ferguson, A.I.

Fraley, R.

R. Fraley, S. Subramani, P. Berg, and D. Papahadjopolous, “Introduction of liposome-encapsulated SV40 DNA into cells,” J. Biol. Chem. 255, 10431–10435 (1980).
[PubMed]

Galun, E.

E. Zeira, A. Manevitch, A. Khatchatouriants, O. Pappo, E. Hyam, M. Darash-Yahana, E. Tavor, A. Honigman, A. Lewis, and E. Galun, “Femtosecond Infrared Laser - An Efficient and Safe in Vivo Gene Delivery System for Prolonged Expression,” Mol. Therapy 8, 342–350 (2003).
[CrossRef]

Gertler, F.B.

D.A. Rubinson, C.P. Dillon, A.V. Kwiatkowski, C. Sievers, L. Yang, J. Kopinja, M. Zhang, M.T. McManus, F.B. Gertler, M.L. Scott, and L. Van Parijs, “A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference,” Nature Genet. 33, 401–406 (2003).
[CrossRef] [PubMed]

Girkin, J.M.

Graham, F.L.

F.L. Graham and A.J. Van der Eb, “A new technique for the assay of infectivity of human adenovirus 5 DNA,” Virology 52, 456–467 (1973).
[CrossRef] [PubMed]

Gunn-Moore, F.

Gupta, P.K.

S.K. Mohanty, M. Sharma, and P.K. Gupta, “Laser-assisted microinjection into targeted animal cells,” Biotechnol. Lett. 25, 895–899 (2003).
[CrossRef] [PubMed]

Hendinger, A.

H. Schneckenburger, A. Hendinger, R. Sailer, W.S.L. Strauss, and M. Schmitt, “Laser-assisted optoporation of single cells,” J. Biomed. Opt. 7, 410–416 (2002).
[CrossRef] [PubMed]

Honigman, A.

E. Zeira, A. Manevitch, A. Khatchatouriants, O. Pappo, E. Hyam, M. Darash-Yahana, E. Tavor, A. Honigman, A. Lewis, and E. Galun, “Femtosecond Infrared Laser - An Efficient and Safe in Vivo Gene Delivery System for Prolonged Expression,” Mol. Therapy 8, 342–350 (2003).
[CrossRef]

Hyam, E.

E. Zeira, A. Manevitch, A. Khatchatouriants, O. Pappo, E. Hyam, M. Darash-Yahana, E. Tavor, A. Honigman, A. Lewis, and E. Galun, “Femtosecond Infrared Laser - An Efficient and Safe in Vivo Gene Delivery System for Prolonged Expression,” Mol. Therapy 8, 342–350 (2003).
[CrossRef]

Itagak, H.

Y. Shirahata, N. Ohkohchi, H. Itagak, and S. Satomi, “New technique for gene transfection using laser irradiation,” J. Invest. Med. 49, 184–190 (2001).
[CrossRef]

Jacobson, S.C.

J.S. Soughayer, T. Krasieva, S.C. Jacobson, J.M. Ramsey, B.J. Tromberg, and N.L. Allbritton, “Characterization of cellular optoporation with distance,” Anal. Chem. 72, 1342–1347 (2000).
[CrossRef] [PubMed]

Khatchatouriants, A.

E. Zeira, A. Manevitch, A. Khatchatouriants, O. Pappo, E. Hyam, M. Darash-Yahana, E. Tavor, A. Honigman, A. Lewis, and E. Galun, “Femtosecond Infrared Laser - An Efficient and Safe in Vivo Gene Delivery System for Prolonged Expression,” Mol. Therapy 8, 342–350 (2003).
[CrossRef]

Konig, K.

U.K. Tirlapur and K. Konig, “Targeted transfection by femtosecond laser,” Nature 418, 290–291 (2002).
[CrossRef] [PubMed]

U.K. Tirlapur and K. Konig, “Femtosecond near-infrared laser pulses as a versatile non- invasive tool for intra-tissue nanoprocessing in plants without compromising viability,” Plant J. 31, 365–374 (2002).
[CrossRef] [PubMed]

Kopinja, J.

D.A. Rubinson, C.P. Dillon, A.V. Kwiatkowski, C. Sievers, L. Yang, J. Kopinja, M. Zhang, M.T. McManus, F.B. Gertler, M.L. Scott, and L. Van Parijs, “A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference,” Nature Genet. 33, 401–406 (2003).
[CrossRef] [PubMed]

Krasieva, T.

J.S. Soughayer, T. Krasieva, S.C. Jacobson, J.M. Ramsey, B.J. Tromberg, and N.L. Allbritton, “Characterization of cellular optoporation with distance,” Anal. Chem. 72, 1342–1347 (2000).
[CrossRef] [PubMed]

Kwiatkowski, A.V.

D.A. Rubinson, C.P. Dillon, A.V. Kwiatkowski, C. Sievers, L. Yang, J. Kopinja, M. Zhang, M.T. McManus, F.B. Gertler, M.L. Scott, and L. Van Parijs, “A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference,” Nature Genet. 33, 401–406 (2003).
[CrossRef] [PubMed]

Lake, T.K.

Lewis, A.

E. Zeira, A. Manevitch, A. Khatchatouriants, O. Pappo, E. Hyam, M. Darash-Yahana, E. Tavor, A. Honigman, A. Lewis, and E. Galun, “Femtosecond Infrared Laser - An Efficient and Safe in Vivo Gene Delivery System for Prolonged Expression,” Mol. Therapy 8, 342–350 (2003).
[CrossRef]

Liu, Y

Y Liu, D. K. Cheng, G. J. Sonek, M. W. Berns, C. F. Chapman, and B J. Tromberg, “Evidence for localized cell heating induced by infrared optical tweezers,” Biophys. J. 68, 2137–2144 (1995).
[CrossRef] [PubMed]

Manevitch, A.

E. Zeira, A. Manevitch, A. Khatchatouriants, O. Pappo, E. Hyam, M. Darash-Yahana, E. Tavor, A. Honigman, A. Lewis, and E. Galun, “Femtosecond Infrared Laser - An Efficient and Safe in Vivo Gene Delivery System for Prolonged Expression,” Mol. Therapy 8, 342–350 (2003).
[CrossRef]

McGloin, D.

D. McGloin and K. Dholakia, private communication

McManus, M.T.

D.A. Rubinson, C.P. Dillon, A.V. Kwiatkowski, C. Sievers, L. Yang, J. Kopinja, M. Zhang, M.T. McManus, F.B. Gertler, M.L. Scott, and L. Van Parijs, “A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference,” Nature Genet. 33, 401–406 (2003).
[CrossRef] [PubMed]

Mohanty, S.K.

S.K. Mohanty, M. Sharma, and P.K. Gupta, “Laser-assisted microinjection into targeted animal cells,” Biotechnol. Lett. 25, 895–899 (2003).
[CrossRef] [PubMed]

Neumann, E.

T.K. Wong and E. Neumann, “Electric-field mediated gene-transfer,” Biochem. and Biophys. Res. Commun. 107, 584–587 (1982).
[CrossRef]

Ohkohchi, N.

Y. Shirahata, N. Ohkohchi, H. Itagak, and S. Satomi, “New technique for gene transfection using laser irradiation,” J. Invest. Med. 49, 184–190 (2001).
[CrossRef]

Palumbo, G.

G. Palumbo, M. Caruso, E. Crescenzi, M.F. Tecce, G. Roberti, and A. Colasanti, “Targeted gene transfer in eucaryotic cells by dye-assisted laser optoporation,” J. Photochem. Photobiol. B-Biol. 36, 41–46 (1996).
[CrossRef]

Papahadjopolous, D.

R. Fraley, S. Subramani, P. Berg, and D. Papahadjopolous, “Introduction of liposome-encapsulated SV40 DNA into cells,” J. Biol. Chem. 255, 10431–10435 (1980).
[PubMed]

Pappo, O.

E. Zeira, A. Manevitch, A. Khatchatouriants, O. Pappo, E. Hyam, M. Darash-Yahana, E. Tavor, A. Honigman, A. Lewis, and E. Galun, “Femtosecond Infrared Laser - An Efficient and Safe in Vivo Gene Delivery System for Prolonged Expression,” Mol. Therapy 8, 342–350 (2003).
[CrossRef]

Paterson, L.

Ramsey, J.M.

J.S. Soughayer, T. Krasieva, S.C. Jacobson, J.M. Ramsey, B.J. Tromberg, and N.L. Allbritton, “Characterization of cellular optoporation with distance,” Anal. Chem. 72, 1342–1347 (2000).
[CrossRef] [PubMed]

Roberti, G.

G. Palumbo, M. Caruso, E. Crescenzi, M.F. Tecce, G. Roberti, and A. Colasanti, “Targeted gene transfer in eucaryotic cells by dye-assisted laser optoporation,” J. Photochem. Photobiol. B-Biol. 36, 41–46 (1996).
[CrossRef]

Rubinson, D.A.

D.A. Rubinson, C.P. Dillon, A.V. Kwiatkowski, C. Sievers, L. Yang, J. Kopinja, M. Zhang, M.T. McManus, F.B. Gertler, M.L. Scott, and L. Van Parijs, “A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference,” Nature Genet. 33, 401–406 (2003).
[CrossRef] [PubMed]

Sailer, R.

H. Schneckenburger, A. Hendinger, R. Sailer, W.S.L. Strauss, and M. Schmitt, “Laser-assisted optoporation of single cells,” J. Biomed. Opt. 7, 410–416 (2002).
[CrossRef] [PubMed]

Satomi, S.

Y. Shirahata, N. Ohkohchi, H. Itagak, and S. Satomi, “New technique for gene transfection using laser irradiation,” J. Invest. Med. 49, 184–190 (2001).
[CrossRef]

Schmitt, M.

H. Schneckenburger, A. Hendinger, R. Sailer, W.S.L. Strauss, and M. Schmitt, “Laser-assisted optoporation of single cells,” J. Biomed. Opt. 7, 410–416 (2002).
[CrossRef] [PubMed]

Schneckenburger, H.

H. Schneckenburger, A. Hendinger, R. Sailer, W.S.L. Strauss, and M. Schmitt, “Laser-assisted optoporation of single cells,” J. Biomed. Opt. 7, 410–416 (2002).
[CrossRef] [PubMed]

Scott, M.L.

D.A. Rubinson, C.P. Dillon, A.V. Kwiatkowski, C. Sievers, L. Yang, J. Kopinja, M. Zhang, M.T. McManus, F.B. Gertler, M.L. Scott, and L. Van Parijs, “A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference,” Nature Genet. 33, 401–406 (2003).
[CrossRef] [PubMed]

Sharma, M.

S.K. Mohanty, M. Sharma, and P.K. Gupta, “Laser-assisted microinjection into targeted animal cells,” Biotechnol. Lett. 25, 895–899 (2003).
[CrossRef] [PubMed]

Shirahata, Y.

Y. Shirahata, N. Ohkohchi, H. Itagak, and S. Satomi, “New technique for gene transfection using laser irradiation,” J. Invest. Med. 49, 184–190 (2001).
[CrossRef]

Sibbett, W.

Sievers, C.

D.A. Rubinson, C.P. Dillon, A.V. Kwiatkowski, C. Sievers, L. Yang, J. Kopinja, M. Zhang, M.T. McManus, F.B. Gertler, M.L. Scott, and L. Van Parijs, “A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference,” Nature Genet. 33, 401–406 (2003).
[CrossRef] [PubMed]

Sonek, G. J.

Y Liu, D. K. Cheng, G. J. Sonek, M. W. Berns, C. F. Chapman, and B J. Tromberg, “Evidence for localized cell heating induced by infrared optical tweezers,” Biophys. J. 68, 2137–2144 (1995).
[CrossRef] [PubMed]

Soughayer, J.S.

J.S. Soughayer, T. Krasieva, S.C. Jacobson, J.M. Ramsey, B.J. Tromberg, and N.L. Allbritton, “Characterization of cellular optoporation with distance,” Anal. Chem. 72, 1342–1347 (2000).
[CrossRef] [PubMed]

Southern, P.

P. Southern and P. Berg, “Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter,” J. Mol. Appl. Genet. 1, 327–341 (1982).
[PubMed]

Strauss, W.S.L.

H. Schneckenburger, A. Hendinger, R. Sailer, W.S.L. Strauss, and M. Schmitt, “Laser-assisted optoporation of single cells,” J. Biomed. Opt. 7, 410–416 (2002).
[CrossRef] [PubMed]

Subramani, S.

R. Fraley, S. Subramani, P. Berg, and D. Papahadjopolous, “Introduction of liposome-encapsulated SV40 DNA into cells,” J. Biol. Chem. 255, 10431–10435 (1980).
[PubMed]

Tavor, E.

E. Zeira, A. Manevitch, A. Khatchatouriants, O. Pappo, E. Hyam, M. Darash-Yahana, E. Tavor, A. Honigman, A. Lewis, and E. Galun, “Femtosecond Infrared Laser - An Efficient and Safe in Vivo Gene Delivery System for Prolonged Expression,” Mol. Therapy 8, 342–350 (2003).
[CrossRef]

Taylor, M.

Tecce, M.F.

G. Palumbo, M. Caruso, E. Crescenzi, M.F. Tecce, G. Roberti, and A. Colasanti, “Targeted gene transfer in eucaryotic cells by dye-assisted laser optoporation,” J. Photochem. Photobiol. B-Biol. 36, 41–46 (1996).
[CrossRef]

Tirlapur, U.K.

U.K. Tirlapur and K. Konig, “Targeted transfection by femtosecond laser,” Nature 418, 290–291 (2002).
[CrossRef] [PubMed]

U.K. Tirlapur and K. Konig, “Femtosecond near-infrared laser pulses as a versatile non- invasive tool for intra-tissue nanoprocessing in plants without compromising viability,” Plant J. 31, 365–374 (2002).
[CrossRef] [PubMed]

Tromberg, B J.

Y Liu, D. K. Cheng, G. J. Sonek, M. W. Berns, C. F. Chapman, and B J. Tromberg, “Evidence for localized cell heating induced by infrared optical tweezers,” Biophys. J. 68, 2137–2144 (1995).
[CrossRef] [PubMed]

Tromberg, B.J.

J.S. Soughayer, T. Krasieva, S.C. Jacobson, J.M. Ramsey, B.J. Tromberg, and N.L. Allbritton, “Characterization of cellular optoporation with distance,” Anal. Chem. 72, 1342–1347 (2000).
[CrossRef] [PubMed]

Van der Eb, A.J.

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Other (1)

D. McGloin and K. Dholakia, private communication

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Figures (4)

Fig. 1.
Fig. 1.

Photoporation apparatus. A violet diode laser (Toptica Photonics, 405 nm, 40 mW output power) is directed towards the sample through an inverted optical microscope setup.

Fig. 2.
Fig. 2.

Captured image of a cell being exposed to the focused violet diode laser beam (0.3 mW average power for 40 milliseconds).

Fig. 3.
Fig. 3.

Fluorescence images of cells transfected with an antibiotic-resistance gene and the GFP gene, taken several weeks after photoporation (images taken using FITC filter on a Zeiss Axioplan 2 imaging microscope). Cells are those that were subcultured from the antibiotic resistant colonies. GFP expression throughout the cells is clear. (a) Live cells (×20); (b) Fixed cells in Vectashield (×100).

Fig. 4.
Fig. 4.

Fluorescence images of live cells transfected with a plasmid containing an antibiotic-resistance gene and a gene encoding a red fluorescent protein (pDsRed-Mito, BD Biosciences, Oxford, U.K.), taken several weeks after photoporation. Expression of the red fluorescent protein in the cell mitochondria is clear. (a) ×20 magnification; (b) ×100 magnification

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