Abstract

Subcellular organelles in living cells were inactivated by tightly focusing femtosecond laser pulses inside the cells. Photodisruption of a mitochondrion in living cells was experimentally confirmed by stacking three-dimensional confocal images and by restaining of organelles. The viability of the cells after femtosecond laser irradiation was ascertained by impermeability of propidium iodide as well as by the presence of cytoplasmic streaming.

© 2004 Optical Society of America

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2004 (3)

N. Shen, M. Colvin, F. Genin, T. Huser, G. A. Cortopassi, T. Stearns, P. LeDuc, D. E. Ingber, and E. Mazur, “Using femtosecond laser subcellular surgery to study cell biology,” Biophys. J. 86, 520A–520A, Part 2 Suppl. S (2004).

T. Higashi, E. Nagamori, T. Sone, S. Matsunaga, and K. Fukui, “A novel transfection method for mammalian cells using calcium alginate microbeads,” J. Biosci. Bioeng. 97, 191–195 (2004).

A. P. Joglekar, H. -H. Liu, E. Meyhöfer, G. Mourou, and A. J. Hunt, Poc. Natl. Acad. Sci. USA 101, 5856–5861 (2004).
[CrossRef]

2002 (4)

C. B. Schaffer, N. Nishimura, E. N. Glezer, A. M.-T. Kim, and E. Mazur, “Dynamics of femtosecond laser-induced breakdown in water from femtoseconds to microseconds,” Opt. Express. 10, 196–203 (2002), http://www.opticsexpress.org/abstract.cfm?URI=OPEX-10-3-196.
[CrossRef] [PubMed]

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

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

V. Venygopalan, A. Guerra III, K. Hahen, and A. Vogel, “Role of laser-induced plasma formation in pulse cellular microsurgery and micromaniplation,” Phys. Rev. Lett. 88, 078103 (2002).
[CrossRef]

2001 (3)

N. Shen, C. B. Schaffer, D. Datta, and E. Mazur, “Photodisruption in biological tissues and single cells using femtosecond laser pulses,” Conference on Lasers and Electro-Optics (Baltimore, MD, 2001) 403–404.

U. K. Tirlapur, K. König, C. Peuckert, R. Krieg, and K. Halbhuber, “Femtosecond near-infrared laser pulse elicit generation of reactive oxygen species in mammalian cells leading to apotosis-like death,” Exp. Cell Res. 263, 88–97 (2001).
[CrossRef] [PubMed]

K. König, W. Riemann, and W. Fritzsche, “Nanodissection of human chromosomes with near-infrared femtosecond laser pulses,” Opt. Lett. 26, 819–821(2001).
[CrossRef]

2000 (2)

K. König, “Laser tweezers and multiphoton microscopes in life sciences,” Histochem. Cell Biol. 114, 79–92 (2000).
[PubMed]

H. Oehring, I. Riedmann, P. Fisher, K. -J. Halbhuber, and K. König, “Ultrastructure and reproduction behaviour of single CHO-K1 cells exposed to near infrared femtosecond laser pulses,” Scanning 22, 263–270 (2000).
[CrossRef] [PubMed]

1999 (5)

J. M. Squirell, D. L. Wokosin, J. G. White, and B. D. Bavister, “Long-term two-photon fluorescence imaging of mammalian embryos without compromising vitality,” Nat. Biotechnolo. 17, 763–767 (1999).
[CrossRef]

H. J. Kester, D. Baur, R. Uhl, and S. W. Hell, “Ca2+ fluorescence imaging with pico- and femtosecond two-photon excitation: signal and photodamage,” Biophys. J. 77, 2226–2236 (1999).
[CrossRef]

K. König, T. W. Becker, P. Fischer, I. Riemann, and K. -J. Halbhuber, “Pulse-length dependence of cellular response to intense near-infrared laser pulses in multiphoton microscopes,” Opt. Lett. 24, 113–115 (1999).
[CrossRef]

E. B. Brown, E. S. Wu, W. Zipfel, and W. W. Webb, “Measurement of molecular diffusion in solution by multiphoton fluorescence photobleaching recovery,” Biophys. J. 77, 2837–2849 (1999).
[CrossRef] [PubMed]

J. White and E. H. Stelzer, “Photobleaching GFP reveals protein dynamics inside live cells,” Trends Cell Biol. 9, 61–65 (1999).
[CrossRef] [PubMed]

1998 (1)

M. Lenzner, J. Krüger, S. Sartania, Z. Cheng, Ch. Spielmann, G. Mourou, W. Kautek, and F. Krausz, “Femtosecond optical breakdown in dielectrics,” Phys. Rev. Lett. 80, 4076–4079 (1998).
[CrossRef]

1997 (3)

Y. Sako, A. Sekihata, Y. Yanagisawa, M. Yamamoto, Y. Shimada, K. Ozaki, and A. Kusumi, “Comparison of two-photon excitation laser scanning microscopy with UV-confocal laser scanning microscopy in three-dimensional calcium imaging using the fluorescence indicator Indo-1,” J. Microsc. 185, 9–20 (1997).
[CrossRef] [PubMed]

K. König, P. T. C. So, W. W. Mantulin, B. J. Tromberg, and E. Gratton, “Cellular response to near-infrared femtosecond laser pulses in two-photon microscopes,” Opt. Lett. 22, 135–136 (1997).
[CrossRef] [PubMed]

E. N. Glezer, C. B. Schaffer, N. Nishimura, and E. Mazur, “Minimally disruptive laser-induced breakdown in water,” Opt. Lett. 22, 1817–1879 (1997).
[CrossRef]

1996 (1)

1991 (1)

M. W. Berns, W. H. Write, and R. W. Steubing, “Laser microbeam as a tool in cell biology,” Int. Rev. Cytol. 129, 1–44 (1991).
[CrossRef] [PubMed]

1990 (1)

W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248, 73–76 (1990).
[CrossRef] [PubMed]

1989 (1)

B. Zysset, J. G. Fujimoto, and T. F. Deutsch, ”Time-resolved measurement of picosecond optical-breakdown,” Appl. Phys. B 48, 139–147(1989).
[CrossRef]

1986 (1)

N. Benvenisty and L. Reshef, “Direct introduction of genes into rats and expression of the genes,” Proc. Natl. Acad. Sci. USA 83, 9551–9555 (1986).
[CrossRef] [PubMed]

1981 (1)

M. W. Berns, J. Aist, J. Edwards, K. Strahs, J. Girton, P. McNeil, J. B. Rattner, M. Kitzes, M. Hammerwilson, L. H. Liaw, A. Siemens, M. Koonce, S. Peterson, S. Brenner, J. Burt, R. Walter, P. J. Bryant, D. Vandyk, J. Couclombe, T. Cahill, and G. S. Bern, “Laser microsurgery in cell and developmental biology,” Science 213, 505–513 (1981).
[CrossRef] [PubMed]

1968 (1)

P. A. Barnes and K. E. Rieckoff, “Laser induced underwater sparks,” Appl. Phys. Lett. 13, 282–284 (1968).
[CrossRef]

1965 (1)

R. L. Amy and R. Storb, “Selective mitochondrial damage by a ruby laser microbeam: an electron microscope study,” Science 150, 756–757 (1965).
[CrossRef] [PubMed]

Aist, J.

M. W. Berns, J. Aist, J. Edwards, K. Strahs, J. Girton, P. McNeil, J. B. Rattner, M. Kitzes, M. Hammerwilson, L. H. Liaw, A. Siemens, M. Koonce, S. Peterson, S. Brenner, J. Burt, R. Walter, P. J. Bryant, D. Vandyk, J. Couclombe, T. Cahill, and G. S. Bern, “Laser microsurgery in cell and developmental biology,” Science 213, 505–513 (1981).
[CrossRef] [PubMed]

Alberts, B.

B. Alberts, D. Bray, A. Johnson, J. Lewis, M. Raff, K. Roberts, and P. Walter, Essential Cell Biology (Taylor and Francis, New York, 1997).

Amy, R. L.

R. L. Amy and R. Storb, “Selective mitochondrial damage by a ruby laser microbeam: an electron microscope study,” Science 150, 756–757 (1965).
[CrossRef] [PubMed]

Barnes, P. A.

P. A. Barnes and K. E. Rieckoff, “Laser induced underwater sparks,” Appl. Phys. Lett. 13, 282–284 (1968).
[CrossRef]

Baur, D.

H. J. Kester, D. Baur, R. Uhl, and S. W. Hell, “Ca2+ fluorescence imaging with pico- and femtosecond two-photon excitation: signal and photodamage,” Biophys. J. 77, 2226–2236 (1999).
[CrossRef]

Bavister, B. D.

J. M. Squirell, D. L. Wokosin, J. G. White, and B. D. Bavister, “Long-term two-photon fluorescence imaging of mammalian embryos without compromising vitality,” Nat. Biotechnolo. 17, 763–767 (1999).
[CrossRef]

Becker, T. W.

Benvenisty, N.

N. Benvenisty and L. Reshef, “Direct introduction of genes into rats and expression of the genes,” Proc. Natl. Acad. Sci. USA 83, 9551–9555 (1986).
[CrossRef] [PubMed]

Bern, G. S.

M. W. Berns, J. Aist, J. Edwards, K. Strahs, J. Girton, P. McNeil, J. B. Rattner, M. Kitzes, M. Hammerwilson, L. H. Liaw, A. Siemens, M. Koonce, S. Peterson, S. Brenner, J. Burt, R. Walter, P. J. Bryant, D. Vandyk, J. Couclombe, T. Cahill, and G. S. Bern, “Laser microsurgery in cell and developmental biology,” Science 213, 505–513 (1981).
[CrossRef] [PubMed]

Berns, M. W.

M. W. Berns, W. H. Write, and R. W. Steubing, “Laser microbeam as a tool in cell biology,” Int. Rev. Cytol. 129, 1–44 (1991).
[CrossRef] [PubMed]

M. W. Berns, J. Aist, J. Edwards, K. Strahs, J. Girton, P. McNeil, J. B. Rattner, M. Kitzes, M. Hammerwilson, L. H. Liaw, A. Siemens, M. Koonce, S. Peterson, S. Brenner, J. Burt, R. Walter, P. J. Bryant, D. Vandyk, J. Couclombe, T. Cahill, and G. S. Bern, “Laser microsurgery in cell and developmental biology,” Science 213, 505–513 (1981).
[CrossRef] [PubMed]

Bray, D.

B. Alberts, D. Bray, A. Johnson, J. Lewis, M. Raff, K. Roberts, and P. Walter, Essential Cell Biology (Taylor and Francis, New York, 1997).

Brenner, S.

M. W. Berns, J. Aist, J. Edwards, K. Strahs, J. Girton, P. McNeil, J. B. Rattner, M. Kitzes, M. Hammerwilson, L. H. Liaw, A. Siemens, M. Koonce, S. Peterson, S. Brenner, J. Burt, R. Walter, P. J. Bryant, D. Vandyk, J. Couclombe, T. Cahill, and G. S. Bern, “Laser microsurgery in cell and developmental biology,” Science 213, 505–513 (1981).
[CrossRef] [PubMed]

Brown, E. B.

E. B. Brown, E. S. Wu, W. Zipfel, and W. W. Webb, “Measurement of molecular diffusion in solution by multiphoton fluorescence photobleaching recovery,” Biophys. J. 77, 2837–2849 (1999).
[CrossRef] [PubMed]

Bryant, P. J.

M. W. Berns, J. Aist, J. Edwards, K. Strahs, J. Girton, P. McNeil, J. B. Rattner, M. Kitzes, M. Hammerwilson, L. H. Liaw, A. Siemens, M. Koonce, S. Peterson, S. Brenner, J. Burt, R. Walter, P. J. Bryant, D. Vandyk, J. Couclombe, T. Cahill, and G. S. Bern, “Laser microsurgery in cell and developmental biology,” Science 213, 505–513 (1981).
[CrossRef] [PubMed]

Burt, J.

M. W. Berns, J. Aist, J. Edwards, K. Strahs, J. Girton, P. McNeil, J. B. Rattner, M. Kitzes, M. Hammerwilson, L. H. Liaw, A. Siemens, M. Koonce, S. Peterson, S. Brenner, J. Burt, R. Walter, P. J. Bryant, D. Vandyk, J. Couclombe, T. Cahill, and G. S. Bern, “Laser microsurgery in cell and developmental biology,” Science 213, 505–513 (1981).
[CrossRef] [PubMed]

Cahill, T.

M. W. Berns, J. Aist, J. Edwards, K. Strahs, J. Girton, P. McNeil, J. B. Rattner, M. Kitzes, M. Hammerwilson, L. H. Liaw, A. Siemens, M. Koonce, S. Peterson, S. Brenner, J. Burt, R. Walter, P. J. Bryant, D. Vandyk, J. Couclombe, T. Cahill, and G. S. Bern, “Laser microsurgery in cell and developmental biology,” Science 213, 505–513 (1981).
[CrossRef] [PubMed]

Cheng, Z.

M. Lenzner, J. Krüger, S. Sartania, Z. Cheng, Ch. Spielmann, G. Mourou, W. Kautek, and F. Krausz, “Femtosecond optical breakdown in dielectrics,” Phys. Rev. Lett. 80, 4076–4079 (1998).
[CrossRef]

Colvin, M.

N. Shen, M. Colvin, F. Genin, T. Huser, G. A. Cortopassi, T. Stearns, P. LeDuc, D. E. Ingber, and E. Mazur, “Using femtosecond laser subcellular surgery to study cell biology,” Biophys. J. 86, 520A–520A, Part 2 Suppl. S (2004).

Cortopassi, G. A.

N. Shen, M. Colvin, F. Genin, T. Huser, G. A. Cortopassi, T. Stearns, P. LeDuc, D. E. Ingber, and E. Mazur, “Using femtosecond laser subcellular surgery to study cell biology,” Biophys. J. 86, 520A–520A, Part 2 Suppl. S (2004).

Couclombe, J.

M. W. Berns, J. Aist, J. Edwards, K. Strahs, J. Girton, P. McNeil, J. B. Rattner, M. Kitzes, M. Hammerwilson, L. H. Liaw, A. Siemens, M. Koonce, S. Peterson, S. Brenner, J. Burt, R. Walter, P. J. Bryant, D. Vandyk, J. Couclombe, T. Cahill, and G. S. Bern, “Laser microsurgery in cell and developmental biology,” Science 213, 505–513 (1981).
[CrossRef] [PubMed]

Datta, D.

N. Shen, C. B. Schaffer, D. Datta, and E. Mazur, “Photodisruption in biological tissues and single cells using femtosecond laser pulses,” Conference on Lasers and Electro-Optics (Baltimore, MD, 2001) 403–404.

Denk, W.

W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248, 73–76 (1990).
[CrossRef] [PubMed]

Deutsch, T. F.

B. Zysset, J. G. Fujimoto, and T. F. Deutsch, ”Time-resolved measurement of picosecond optical-breakdown,” Appl. Phys. B 48, 139–147(1989).
[CrossRef]

Edwards, J.

M. W. Berns, J. Aist, J. Edwards, K. Strahs, J. Girton, P. McNeil, J. B. Rattner, M. Kitzes, M. Hammerwilson, L. H. Liaw, A. Siemens, M. Koonce, S. Peterson, S. Brenner, J. Burt, R. Walter, P. J. Bryant, D. Vandyk, J. Couclombe, T. Cahill, and G. S. Bern, “Laser microsurgery in cell and developmental biology,” Science 213, 505–513 (1981).
[CrossRef] [PubMed]

Feit, M. D.

Fischer, P.

Fisher, P.

H. Oehring, I. Riedmann, P. Fisher, K. -J. Halbhuber, and K. König, “Ultrastructure and reproduction behaviour of single CHO-K1 cells exposed to near infrared femtosecond laser pulses,” Scanning 22, 263–270 (2000).
[CrossRef] [PubMed]

Fritzsche, W.

Fujimoto, J. G.

B. Zysset, J. G. Fujimoto, and T. F. Deutsch, ”Time-resolved measurement of picosecond optical-breakdown,” Appl. Phys. B 48, 139–147(1989).
[CrossRef]

Fukui, K.

T. Higashi, E. Nagamori, T. Sone, S. Matsunaga, and K. Fukui, “A novel transfection method for mammalian cells using calcium alginate microbeads,” J. Biosci. Bioeng. 97, 191–195 (2004).

Genin, F.

N. Shen, M. Colvin, F. Genin, T. Huser, G. A. Cortopassi, T. Stearns, P. LeDuc, D. E. Ingber, and E. Mazur, “Using femtosecond laser subcellular surgery to study cell biology,” Biophys. J. 86, 520A–520A, Part 2 Suppl. S (2004).

Girton, J.

M. W. Berns, J. Aist, J. Edwards, K. Strahs, J. Girton, P. McNeil, J. B. Rattner, M. Kitzes, M. Hammerwilson, L. H. Liaw, A. Siemens, M. Koonce, S. Peterson, S. Brenner, J. Burt, R. Walter, P. J. Bryant, D. Vandyk, J. Couclombe, T. Cahill, and G. S. Bern, “Laser microsurgery in cell and developmental biology,” Science 213, 505–513 (1981).
[CrossRef] [PubMed]

Glezer, E. N.

C. B. Schaffer, N. Nishimura, E. N. Glezer, A. M.-T. Kim, and E. Mazur, “Dynamics of femtosecond laser-induced breakdown in water from femtoseconds to microseconds,” Opt. Express. 10, 196–203 (2002), http://www.opticsexpress.org/abstract.cfm?URI=OPEX-10-3-196.
[CrossRef] [PubMed]

E. N. Glezer, C. B. Schaffer, N. Nishimura, and E. Mazur, “Minimally disruptive laser-induced breakdown in water,” Opt. Lett. 22, 1817–1879 (1997).
[CrossRef]

Gratton, E.

Greulich, K. O.

K. O. Greulich, Micromanipulation by light in biology and medicine, (Birkhähser-Verlag, Basel, Switzerland, 1999).

Guerra III, A.

V. Venygopalan, A. Guerra III, K. Hahen, and A. Vogel, “Role of laser-induced plasma formation in pulse cellular microsurgery and micromaniplation,” Phys. Rev. Lett. 88, 078103 (2002).
[CrossRef]

Hahen, K.

V. Venygopalan, A. Guerra III, K. Hahen, and A. Vogel, “Role of laser-induced plasma formation in pulse cellular microsurgery and micromaniplation,” Phys. Rev. Lett. 88, 078103 (2002).
[CrossRef]

Halbhuber, K.

U. K. Tirlapur, K. König, C. Peuckert, R. Krieg, and K. Halbhuber, “Femtosecond near-infrared laser pulse elicit generation of reactive oxygen species in mammalian cells leading to apotosis-like death,” Exp. Cell Res. 263, 88–97 (2001).
[CrossRef] [PubMed]

Halbhuber, K. -J.

H. Oehring, I. Riedmann, P. Fisher, K. -J. Halbhuber, and K. König, “Ultrastructure and reproduction behaviour of single CHO-K1 cells exposed to near infrared femtosecond laser pulses,” Scanning 22, 263–270 (2000).
[CrossRef] [PubMed]

K. König, T. W. Becker, P. Fischer, I. Riemann, and K. -J. Halbhuber, “Pulse-length dependence of cellular response to intense near-infrared laser pulses in multiphoton microscopes,” Opt. Lett. 24, 113–115 (1999).
[CrossRef]

Hammerwilson, M.

M. W. Berns, J. Aist, J. Edwards, K. Strahs, J. Girton, P. McNeil, J. B. Rattner, M. Kitzes, M. Hammerwilson, L. H. Liaw, A. Siemens, M. Koonce, S. Peterson, S. Brenner, J. Burt, R. Walter, P. J. Bryant, D. Vandyk, J. Couclombe, T. Cahill, and G. S. Bern, “Laser microsurgery in cell and developmental biology,” Science 213, 505–513 (1981).
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H. J. Kester, D. Baur, R. Uhl, and S. W. Hell, “Ca2+ fluorescence imaging with pico- and femtosecond two-photon excitation: signal and photodamage,” Biophys. J. 77, 2226–2236 (1999).
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T. Higashi, E. Nagamori, T. Sone, S. Matsunaga, and K. Fukui, “A novel transfection method for mammalian cells using calcium alginate microbeads,” J. Biosci. Bioeng. 97, 191–195 (2004).

Hunt, A. J.

A. P. Joglekar, H. -H. Liu, E. Meyhöfer, G. Mourou, and A. J. Hunt, Poc. Natl. Acad. Sci. USA 101, 5856–5861 (2004).
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Huser, T.

N. Shen, M. Colvin, F. Genin, T. Huser, G. A. Cortopassi, T. Stearns, P. LeDuc, D. E. Ingber, and E. Mazur, “Using femtosecond laser subcellular surgery to study cell biology,” Biophys. J. 86, 520A–520A, Part 2 Suppl. S (2004).

Ingber, D. E.

N. Shen, M. Colvin, F. Genin, T. Huser, G. A. Cortopassi, T. Stearns, P. LeDuc, D. E. Ingber, and E. Mazur, “Using femtosecond laser subcellular surgery to study cell biology,” Biophys. J. 86, 520A–520A, Part 2 Suppl. S (2004).

Joglekar, A. P.

A. P. Joglekar, H. -H. Liu, E. Meyhöfer, G. Mourou, and A. J. Hunt, Poc. Natl. Acad. Sci. USA 101, 5856–5861 (2004).
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B. Alberts, D. Bray, A. Johnson, J. Lewis, M. Raff, K. Roberts, and P. Walter, Essential Cell Biology (Taylor and Francis, New York, 1997).

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M. Lenzner, J. Krüger, S. Sartania, Z. Cheng, Ch. Spielmann, G. Mourou, W. Kautek, and F. Krausz, “Femtosecond optical breakdown in dielectrics,” Phys. Rev. Lett. 80, 4076–4079 (1998).
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H. J. Kester, D. Baur, R. Uhl, and S. W. Hell, “Ca2+ fluorescence imaging with pico- and femtosecond two-photon excitation: signal and photodamage,” Biophys. J. 77, 2226–2236 (1999).
[CrossRef]

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C. B. Schaffer, N. Nishimura, E. N. Glezer, A. M.-T. Kim, and E. Mazur, “Dynamics of femtosecond laser-induced breakdown in water from femtoseconds to microseconds,” Opt. Express. 10, 196–203 (2002), http://www.opticsexpress.org/abstract.cfm?URI=OPEX-10-3-196.
[CrossRef] [PubMed]

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M. W. Berns, J. Aist, J. Edwards, K. Strahs, J. Girton, P. McNeil, J. B. Rattner, M. Kitzes, M. Hammerwilson, L. H. Liaw, A. Siemens, M. Koonce, S. Peterson, S. Brenner, J. Burt, R. Walter, P. J. Bryant, D. Vandyk, J. Couclombe, T. Cahill, and G. S. Bern, “Laser microsurgery in cell and developmental biology,” Science 213, 505–513 (1981).
[CrossRef] [PubMed]

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

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

U. K. Tirlapur, K. König, C. Peuckert, R. Krieg, and K. Halbhuber, “Femtosecond near-infrared laser pulse elicit generation of reactive oxygen species in mammalian cells leading to apotosis-like death,” Exp. Cell Res. 263, 88–97 (2001).
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K. König, W. Riemann, and W. Fritzsche, “Nanodissection of human chromosomes with near-infrared femtosecond laser pulses,” Opt. Lett. 26, 819–821(2001).
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K. König, “Laser tweezers and multiphoton microscopes in life sciences,” Histochem. Cell Biol. 114, 79–92 (2000).
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H. Oehring, I. Riedmann, P. Fisher, K. -J. Halbhuber, and K. König, “Ultrastructure and reproduction behaviour of single CHO-K1 cells exposed to near infrared femtosecond laser pulses,” Scanning 22, 263–270 (2000).
[CrossRef] [PubMed]

K. König, T. W. Becker, P. Fischer, I. Riemann, and K. -J. Halbhuber, “Pulse-length dependence of cellular response to intense near-infrared laser pulses in multiphoton microscopes,” Opt. Lett. 24, 113–115 (1999).
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K. König, P. T. C. So, W. W. Mantulin, B. J. Tromberg, and E. Gratton, “Cellular response to near-infrared femtosecond laser pulses in two-photon microscopes,” Opt. Lett. 22, 135–136 (1997).
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M. W. Berns, J. Aist, J. Edwards, K. Strahs, J. Girton, P. McNeil, J. B. Rattner, M. Kitzes, M. Hammerwilson, L. H. Liaw, A. Siemens, M. Koonce, S. Peterson, S. Brenner, J. Burt, R. Walter, P. J. Bryant, D. Vandyk, J. Couclombe, T. Cahill, and G. S. Bern, “Laser microsurgery in cell and developmental biology,” Science 213, 505–513 (1981).
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M. Lenzner, J. Krüger, S. Sartania, Z. Cheng, Ch. Spielmann, G. Mourou, W. Kautek, and F. Krausz, “Femtosecond optical breakdown in dielectrics,” Phys. Rev. Lett. 80, 4076–4079 (1998).
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U. K. Tirlapur, K. König, C. Peuckert, R. Krieg, and K. Halbhuber, “Femtosecond near-infrared laser pulse elicit generation of reactive oxygen species in mammalian cells leading to apotosis-like death,” Exp. Cell Res. 263, 88–97 (2001).
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Krüger, J.

M. Lenzner, J. Krüger, S. Sartania, Z. Cheng, Ch. Spielmann, G. Mourou, W. Kautek, and F. Krausz, “Femtosecond optical breakdown in dielectrics,” Phys. Rev. Lett. 80, 4076–4079 (1998).
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Y. Sako, A. Sekihata, Y. Yanagisawa, M. Yamamoto, Y. Shimada, K. Ozaki, and A. Kusumi, “Comparison of two-photon excitation laser scanning microscopy with UV-confocal laser scanning microscopy in three-dimensional calcium imaging using the fluorescence indicator Indo-1,” J. Microsc. 185, 9–20 (1997).
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N. Shen, M. Colvin, F. Genin, T. Huser, G. A. Cortopassi, T. Stearns, P. LeDuc, D. E. Ingber, and E. Mazur, “Using femtosecond laser subcellular surgery to study cell biology,” Biophys. J. 86, 520A–520A, Part 2 Suppl. S (2004).

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M. Lenzner, J. Krüger, S. Sartania, Z. Cheng, Ch. Spielmann, G. Mourou, W. Kautek, and F. Krausz, “Femtosecond optical breakdown in dielectrics,” Phys. Rev. Lett. 80, 4076–4079 (1998).
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B. Alberts, D. Bray, A. Johnson, J. Lewis, M. Raff, K. Roberts, and P. Walter, Essential Cell Biology (Taylor and Francis, New York, 1997).

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M. W. Berns, J. Aist, J. Edwards, K. Strahs, J. Girton, P. McNeil, J. B. Rattner, M. Kitzes, M. Hammerwilson, L. H. Liaw, A. Siemens, M. Koonce, S. Peterson, S. Brenner, J. Burt, R. Walter, P. J. Bryant, D. Vandyk, J. Couclombe, T. Cahill, and G. S. Bern, “Laser microsurgery in cell and developmental biology,” Science 213, 505–513 (1981).
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A. P. Joglekar, H. -H. Liu, E. Meyhöfer, G. Mourou, and A. J. Hunt, Poc. Natl. Acad. Sci. USA 101, 5856–5861 (2004).
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Mantulin, W. W.

Matsunaga, S.

T. Higashi, E. Nagamori, T. Sone, S. Matsunaga, and K. Fukui, “A novel transfection method for mammalian cells using calcium alginate microbeads,” J. Biosci. Bioeng. 97, 191–195 (2004).

Mazur, E.

N. Shen, M. Colvin, F. Genin, T. Huser, G. A. Cortopassi, T. Stearns, P. LeDuc, D. E. Ingber, and E. Mazur, “Using femtosecond laser subcellular surgery to study cell biology,” Biophys. J. 86, 520A–520A, Part 2 Suppl. S (2004).

C. B. Schaffer, N. Nishimura, E. N. Glezer, A. M.-T. Kim, and E. Mazur, “Dynamics of femtosecond laser-induced breakdown in water from femtoseconds to microseconds,” Opt. Express. 10, 196–203 (2002), http://www.opticsexpress.org/abstract.cfm?URI=OPEX-10-3-196.
[CrossRef] [PubMed]

N. Shen, C. B. Schaffer, D. Datta, and E. Mazur, “Photodisruption in biological tissues and single cells using femtosecond laser pulses,” Conference on Lasers and Electro-Optics (Baltimore, MD, 2001) 403–404.

E. N. Glezer, C. B. Schaffer, N. Nishimura, and E. Mazur, “Minimally disruptive laser-induced breakdown in water,” Opt. Lett. 22, 1817–1879 (1997).
[CrossRef]

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M. W. Berns, J. Aist, J. Edwards, K. Strahs, J. Girton, P. McNeil, J. B. Rattner, M. Kitzes, M. Hammerwilson, L. H. Liaw, A. Siemens, M. Koonce, S. Peterson, S. Brenner, J. Burt, R. Walter, P. J. Bryant, D. Vandyk, J. Couclombe, T. Cahill, and G. S. Bern, “Laser microsurgery in cell and developmental biology,” Science 213, 505–513 (1981).
[CrossRef] [PubMed]

Meyhöfer, E.

A. P. Joglekar, H. -H. Liu, E. Meyhöfer, G. Mourou, and A. J. Hunt, Poc. Natl. Acad. Sci. USA 101, 5856–5861 (2004).
[CrossRef]

Mourou, G.

A. P. Joglekar, H. -H. Liu, E. Meyhöfer, G. Mourou, and A. J. Hunt, Poc. Natl. Acad. Sci. USA 101, 5856–5861 (2004).
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M. Lenzner, J. Krüger, S. Sartania, Z. Cheng, Ch. Spielmann, G. Mourou, W. Kautek, and F. Krausz, “Femtosecond optical breakdown in dielectrics,” Phys. Rev. Lett. 80, 4076–4079 (1998).
[CrossRef]

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T. Higashi, E. Nagamori, T. Sone, S. Matsunaga, and K. Fukui, “A novel transfection method for mammalian cells using calcium alginate microbeads,” J. Biosci. Bioeng. 97, 191–195 (2004).

Nishimura, N.

C. B. Schaffer, N. Nishimura, E. N. Glezer, A. M.-T. Kim, and E. Mazur, “Dynamics of femtosecond laser-induced breakdown in water from femtoseconds to microseconds,” Opt. Express. 10, 196–203 (2002), http://www.opticsexpress.org/abstract.cfm?URI=OPEX-10-3-196.
[CrossRef] [PubMed]

E. N. Glezer, C. B. Schaffer, N. Nishimura, and E. Mazur, “Minimally disruptive laser-induced breakdown in water,” Opt. Lett. 22, 1817–1879 (1997).
[CrossRef]

Oehring, H.

H. Oehring, I. Riedmann, P. Fisher, K. -J. Halbhuber, and K. König, “Ultrastructure and reproduction behaviour of single CHO-K1 cells exposed to near infrared femtosecond laser pulses,” Scanning 22, 263–270 (2000).
[CrossRef] [PubMed]

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Y. Sako, A. Sekihata, Y. Yanagisawa, M. Yamamoto, Y. Shimada, K. Ozaki, and A. Kusumi, “Comparison of two-photon excitation laser scanning microscopy with UV-confocal laser scanning microscopy in three-dimensional calcium imaging using the fluorescence indicator Indo-1,” J. Microsc. 185, 9–20 (1997).
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Peterson, S.

M. W. Berns, J. Aist, J. Edwards, K. Strahs, J. Girton, P. McNeil, J. B. Rattner, M. Kitzes, M. Hammerwilson, L. H. Liaw, A. Siemens, M. Koonce, S. Peterson, S. Brenner, J. Burt, R. Walter, P. J. Bryant, D. Vandyk, J. Couclombe, T. Cahill, and G. S. Bern, “Laser microsurgery in cell and developmental biology,” Science 213, 505–513 (1981).
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U. K. Tirlapur, K. König, C. Peuckert, R. Krieg, and K. Halbhuber, “Femtosecond near-infrared laser pulse elicit generation of reactive oxygen species in mammalian cells leading to apotosis-like death,” Exp. Cell Res. 263, 88–97 (2001).
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B. Alberts, D. Bray, A. Johnson, J. Lewis, M. Raff, K. Roberts, and P. Walter, Essential Cell Biology (Taylor and Francis, New York, 1997).

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M. W. Berns, J. Aist, J. Edwards, K. Strahs, J. Girton, P. McNeil, J. B. Rattner, M. Kitzes, M. Hammerwilson, L. H. Liaw, A. Siemens, M. Koonce, S. Peterson, S. Brenner, J. Burt, R. Walter, P. J. Bryant, D. Vandyk, J. Couclombe, T. Cahill, and G. S. Bern, “Laser microsurgery in cell and developmental biology,” Science 213, 505–513 (1981).
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H. Oehring, I. Riedmann, P. Fisher, K. -J. Halbhuber, and K. König, “Ultrastructure and reproduction behaviour of single CHO-K1 cells exposed to near infrared femtosecond laser pulses,” Scanning 22, 263–270 (2000).
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Riemann, I.

Riemann, W.

Roberts, K.

B. Alberts, D. Bray, A. Johnson, J. Lewis, M. Raff, K. Roberts, and P. Walter, Essential Cell Biology (Taylor and Francis, New York, 1997).

Rubenchik, A.M.

Sako, Y.

Y. Sako, A. Sekihata, Y. Yanagisawa, M. Yamamoto, Y. Shimada, K. Ozaki, and A. Kusumi, “Comparison of two-photon excitation laser scanning microscopy with UV-confocal laser scanning microscopy in three-dimensional calcium imaging using the fluorescence indicator Indo-1,” J. Microsc. 185, 9–20 (1997).
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Sartania, S.

M. Lenzner, J. Krüger, S. Sartania, Z. Cheng, Ch. Spielmann, G. Mourou, W. Kautek, and F. Krausz, “Femtosecond optical breakdown in dielectrics,” Phys. Rev. Lett. 80, 4076–4079 (1998).
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C. B. Schaffer, N. Nishimura, E. N. Glezer, A. M.-T. Kim, and E. Mazur, “Dynamics of femtosecond laser-induced breakdown in water from femtoseconds to microseconds,” Opt. Express. 10, 196–203 (2002), http://www.opticsexpress.org/abstract.cfm?URI=OPEX-10-3-196.
[CrossRef] [PubMed]

N. Shen, C. B. Schaffer, D. Datta, and E. Mazur, “Photodisruption in biological tissues and single cells using femtosecond laser pulses,” Conference on Lasers and Electro-Optics (Baltimore, MD, 2001) 403–404.

E. N. Glezer, C. B. Schaffer, N. Nishimura, and E. Mazur, “Minimally disruptive laser-induced breakdown in water,” Opt. Lett. 22, 1817–1879 (1997).
[CrossRef]

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Y. Sako, A. Sekihata, Y. Yanagisawa, M. Yamamoto, Y. Shimada, K. Ozaki, and A. Kusumi, “Comparison of two-photon excitation laser scanning microscopy with UV-confocal laser scanning microscopy in three-dimensional calcium imaging using the fluorescence indicator Indo-1,” J. Microsc. 185, 9–20 (1997).
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N. Shen, M. Colvin, F. Genin, T. Huser, G. A. Cortopassi, T. Stearns, P. LeDuc, D. E. Ingber, and E. Mazur, “Using femtosecond laser subcellular surgery to study cell biology,” Biophys. J. 86, 520A–520A, Part 2 Suppl. S (2004).

N. Shen, C. B. Schaffer, D. Datta, and E. Mazur, “Photodisruption in biological tissues and single cells using femtosecond laser pulses,” Conference on Lasers and Electro-Optics (Baltimore, MD, 2001) 403–404.

Shimada, Y.

Y. Sako, A. Sekihata, Y. Yanagisawa, M. Yamamoto, Y. Shimada, K. Ozaki, and A. Kusumi, “Comparison of two-photon excitation laser scanning microscopy with UV-confocal laser scanning microscopy in three-dimensional calcium imaging using the fluorescence indicator Indo-1,” J. Microsc. 185, 9–20 (1997).
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Siemens, A.

M. W. Berns, J. Aist, J. Edwards, K. Strahs, J. Girton, P. McNeil, J. B. Rattner, M. Kitzes, M. Hammerwilson, L. H. Liaw, A. Siemens, M. Koonce, S. Peterson, S. Brenner, J. Burt, R. Walter, P. J. Bryant, D. Vandyk, J. Couclombe, T. Cahill, and G. S. Bern, “Laser microsurgery in cell and developmental biology,” Science 213, 505–513 (1981).
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Sone, T.

T. Higashi, E. Nagamori, T. Sone, S. Matsunaga, and K. Fukui, “A novel transfection method for mammalian cells using calcium alginate microbeads,” J. Biosci. Bioeng. 97, 191–195 (2004).

Spielmann, Ch.

M. Lenzner, J. Krüger, S. Sartania, Z. Cheng, Ch. Spielmann, G. Mourou, W. Kautek, and F. Krausz, “Femtosecond optical breakdown in dielectrics,” Phys. Rev. Lett. 80, 4076–4079 (1998).
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J. M. Squirell, D. L. Wokosin, J. G. White, and B. D. Bavister, “Long-term two-photon fluorescence imaging of mammalian embryos without compromising vitality,” Nat. Biotechnolo. 17, 763–767 (1999).
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N. Shen, M. Colvin, F. Genin, T. Huser, G. A. Cortopassi, T. Stearns, P. LeDuc, D. E. Ingber, and E. Mazur, “Using femtosecond laser subcellular surgery to study cell biology,” Biophys. J. 86, 520A–520A, Part 2 Suppl. S (2004).

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J. White and E. H. Stelzer, “Photobleaching GFP reveals protein dynamics inside live cells,” Trends Cell Biol. 9, 61–65 (1999).
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M. W. Berns, W. H. Write, and R. W. Steubing, “Laser microbeam as a tool in cell biology,” Int. Rev. Cytol. 129, 1–44 (1991).
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M. W. Berns, J. Aist, J. Edwards, K. Strahs, J. Girton, P. McNeil, J. B. Rattner, M. Kitzes, M. Hammerwilson, L. H. Liaw, A. Siemens, M. Koonce, S. Peterson, S. Brenner, J. Burt, R. Walter, P. J. Bryant, D. Vandyk, J. Couclombe, T. Cahill, and G. S. Bern, “Laser microsurgery in cell and developmental biology,” Science 213, 505–513 (1981).
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W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248, 73–76 (1990).
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Tirlapur, U. K.

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

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

U. K. Tirlapur, K. König, C. Peuckert, R. Krieg, and K. Halbhuber, “Femtosecond near-infrared laser pulse elicit generation of reactive oxygen species in mammalian cells leading to apotosis-like death,” Exp. Cell Res. 263, 88–97 (2001).
[CrossRef] [PubMed]

Tromberg, B. J.

Uhl, R.

H. J. Kester, D. Baur, R. Uhl, and S. W. Hell, “Ca2+ fluorescence imaging with pico- and femtosecond two-photon excitation: signal and photodamage,” Biophys. J. 77, 2226–2236 (1999).
[CrossRef]

Vandyk, D.

M. W. Berns, J. Aist, J. Edwards, K. Strahs, J. Girton, P. McNeil, J. B. Rattner, M. Kitzes, M. Hammerwilson, L. H. Liaw, A. Siemens, M. Koonce, S. Peterson, S. Brenner, J. Burt, R. Walter, P. J. Bryant, D. Vandyk, J. Couclombe, T. Cahill, and G. S. Bern, “Laser microsurgery in cell and developmental biology,” Science 213, 505–513 (1981).
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B. Alberts, D. Bray, A. Johnson, J. Lewis, M. Raff, K. Roberts, and P. Walter, Essential Cell Biology (Taylor and Francis, New York, 1997).

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M. W. Berns, J. Aist, J. Edwards, K. Strahs, J. Girton, P. McNeil, J. B. Rattner, M. Kitzes, M. Hammerwilson, L. H. Liaw, A. Siemens, M. Koonce, S. Peterson, S. Brenner, J. Burt, R. Walter, P. J. Bryant, D. Vandyk, J. Couclombe, T. Cahill, and G. S. Bern, “Laser microsurgery in cell and developmental biology,” Science 213, 505–513 (1981).
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E. B. Brown, E. S. Wu, W. Zipfel, and W. W. Webb, “Measurement of molecular diffusion in solution by multiphoton fluorescence photobleaching recovery,” Biophys. J. 77, 2837–2849 (1999).
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J. White and E. H. Stelzer, “Photobleaching GFP reveals protein dynamics inside live cells,” Trends Cell Biol. 9, 61–65 (1999).
[CrossRef] [PubMed]

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J. M. Squirell, D. L. Wokosin, J. G. White, and B. D. Bavister, “Long-term two-photon fluorescence imaging of mammalian embryos without compromising vitality,” Nat. Biotechnolo. 17, 763–767 (1999).
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J. M. Squirell, D. L. Wokosin, J. G. White, and B. D. Bavister, “Long-term two-photon fluorescence imaging of mammalian embryos without compromising vitality,” Nat. Biotechnolo. 17, 763–767 (1999).
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M. W. Berns, W. H. Write, and R. W. Steubing, “Laser microbeam as a tool in cell biology,” Int. Rev. Cytol. 129, 1–44 (1991).
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E. B. Brown, E. S. Wu, W. Zipfel, and W. W. Webb, “Measurement of molecular diffusion in solution by multiphoton fluorescence photobleaching recovery,” Biophys. J. 77, 2837–2849 (1999).
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Y. Sako, A. Sekihata, Y. Yanagisawa, M. Yamamoto, Y. Shimada, K. Ozaki, and A. Kusumi, “Comparison of two-photon excitation laser scanning microscopy with UV-confocal laser scanning microscopy in three-dimensional calcium imaging using the fluorescence indicator Indo-1,” J. Microsc. 185, 9–20 (1997).
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Y. Sako, A. Sekihata, Y. Yanagisawa, M. Yamamoto, Y. Shimada, K. Ozaki, and A. Kusumi, “Comparison of two-photon excitation laser scanning microscopy with UV-confocal laser scanning microscopy in three-dimensional calcium imaging using the fluorescence indicator Indo-1,” J. Microsc. 185, 9–20 (1997).
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E. B. Brown, E. S. Wu, W. Zipfel, and W. W. Webb, “Measurement of molecular diffusion in solution by multiphoton fluorescence photobleaching recovery,” Biophys. J. 77, 2837–2849 (1999).
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Figures (5)

Fig. 1.
Fig. 1.

Schematic of the experimental setup. OB, DM, BP, and PMT denote objective lens, dichroic mirror, bandpass filter, and photomulutiplier tube, respectively.

Fig. 2.
Fig. 2.

Confocal images of the cells (a) before and (b) after laser irradiation. Red fluorescence shows mitochondria of HeLa cells stained with MitoTracker Red. The white circles and arrows indicate individual HeLa cells and target mitochondria, respectively. Additional red fluorescence in Fig. 2b is derived from propidium iodide (PI). The laser pulses were focused inside cells α and β at energies of 7 nJ/pulse and 3 nJ /pulse, respectively.

Fig. 3
Fig. 3

Confocal images of HeLa cell before and after fs-laser irradiation. (a) and (b) confocal images of the HeLa cell before and after femtosecond laser irradiation. Red fluorescence shows mitochondria of HeLa cells stained with MitoTracker Red. A target mitochondrion is indicated by a white arrow. (c) and (d) magnified views of square areas indicated in (a) and (b), respectively. The center of the dotted circles show target mitochondria.

Fig. 4.
Fig. 4.

(a) and (b) stacked three-dimensional confocal images at different depths before and after femtosecond laser irradiation. Red fluorescence shows mitochondria of HeLa cells stained with MitoTracker Red. A target mitochondrion is indicated by a white arrow. Twenty-one images were obtained by translation of the objective lens by 6 μm in steps of 300 nm. (c) and (d) magnified views of square areas indicated in (a) and (b), respectively. The center of the dotted circles show target mitochondria.

Fig. 5.
Fig. 5.

(a) Confocal images of laser-irradiated cells before and after restaining. Yellow fluorescence shows mitochondria visualized by EYFP. Target mitochondrion is indicated by a white arrow. (c) magnified view of square area indicated in (a) before femtosecond laser irradiation. (b) confocal image obtained after femtosecond laser irradiation. (d) magnified view of square area in (b). (e) and (f) confocal images obtained after restaining by MitoTracker Red. (e) confocal image obtained by excitation with the Ar+ laser. (f) confocal image obtained by excitation with the He-Ne laser. Dotted circles show target mitochondria.

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