Abstract

We present a study characterizing the properties of femtosecond laser nanosurgery applied to individual axons in live Caenorhabditis elegans (C. elegans) using nano-Joule laser pulses at 1 kHz repetition rate. Emphasis is placed on the characterization of the damage threshold, the extent of damage, and the statistical rates of axonal recovery as a function of laser parameters. The ablation threshold decreases with increasing number of pulses applied during nanoaxotomy. This dependency suggests the existence of an incubation effect. In terms of extent of damage, the energy per pulse is found to be a more critical parameter than the number of pulses. Axonal recovery improves when surgery is performed using a large number of low energy pulses.

© 2007 Optical Society of America

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  12. A. Heisterkamp, I. Zaharieva Maxwell, E. Mazur, J. M. Underwood, J. A. Nickerson, S. Kumar and D. E. Ingber, "Pulse energy dependence of subcellular dissection by femtosecond laser pulses," Opt. Express 13, 3690-3696 (2005).
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2006

S.H. Chung, D.A. Clark, C.V. Gabel, E. Mazur and A.D.T. Samuel, "The role of the AFD neuron in C. elegans thermotaxis analyzed using femtosecond laser ablation," BMC Neuro. 7:30 (2006).

F. Yanik, H. Cinar, N. Cinar, A. Chisholm, Y. Jin and A. Ben-Yakar, "Nerve regeneration in Caenorhabditis elegans after femtosecond laser axotomy," IEEE J. Sel. Top. Quantum Electron. 12 (6) (2006).

2005

B. Beirowski,R. Adalbert, D. Wagner, D.S. Grumme, K. Addicks, R.R. Ribchester and M.P. Coleman, "The progressive nature of Wallerian degeneration in wild-type and slow Wallerian degeneration (WldS) nerves," BMC Neurosci. 6 (6), (2005).
[CrossRef] [PubMed]

A. Heisterkamp, I. Zaharieva Maxwell, E. Mazur, J. M. Underwood, J. A. Nickerson, S. Kumar and D. E. Ingber, "Pulse energy dependence of subcellular dissection by femtosecond laser pulses," Opt. Express 13, 3690-3696 (2005).
[CrossRef] [PubMed]

N. Shen, D. Datta, C.B. Schaffer, P. LeDuc, D.E. Ingber and E. Mazur, "Ablation of cytoskeletal filaments and mitochondria in cells using a femtosecond laser nanocissor," Mech. Chem. Biosyst. 2, 17 (2005).

M. Kerschensteiner, M.E. Schwab, J.W. Lichtman and T. Misgeld, "In vivo imaging of axonal degereneration and regeneration in the injured spinal cord," Nature Med. 11(5), 572-577 (2005).

A. Vogel, J. Noack, G. Hüttman and G. Paltauf, "Mechanisms of femtosecond laser nanosurgery of cells and tissues," App. Phys. B, 10.1007 (2005).
[CrossRef]

2004

D.H. Bhatt, S.J. Otto, B. Depoister and J.R. Fetcho, "Cyclic AMP-induced repair of zebrafish spinal circuits," Science 305, 254-258 (2004).
[CrossRef] [PubMed]

M.F. Yanik, H. Cinar, H.N. Cinar, A.D. Chisholm, Y. Jin and A. Ben-Yakar, "Functional regeneration after laser axotomy," Nature 432, 882 (2004).
[CrossRef]

H. Urey, "Spot size, depth of focus and diffraction ring intensity formulas for truncated Gaussian beams." App. Phys. 43 (3), 620-625 (2004).

W. Watanabe and N. Arakawa, "Femtosecond laser disruption of subcellular organelles in a living cell," Opt. Express 12 (18), 4203-4213 (2004).
[CrossRef] [PubMed]

2002

X. Huang, H.J. Cheng, M. Tessier-Lavigne, Y. Jin, "MAX-1, a novel PH/MyTH4/FERM domain cytoplasmic protein implicated in netrin-mediated axon repulsion" Neuron 34, 563-576 (2002).
[CrossRef] [PubMed]

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

V. Venugopalan, A. GuerraIII, K. Hahen and A. Vogel, "Role of laser-induced plasma formation in pulse cellular microsurgery and micromanipulation," Phys. Rev. Lett. 88, 078103 (2002).
[CrossRef] [PubMed]

2001

2000

P.J. Horner and F.H. Gage, "Regenerating the damaged central nervous system," Nature 407, 963-970 (2000).
[CrossRef] [PubMed]

1999

M.D. Perry, D.C. Stuart, P.S. Banks, M.D. Feit, V. Yanovsky and A.M. Rubenchik, "Ultrashort-pulse laser machining of dielectric materials," J. Appl. Phys. 85, 6803-6810 (1999).
[CrossRef]

A. Rosenfeld, M. Lorenz, R. Stoian and D. Ashkenasi, "Ultrashort-laser-pulse damage threshold of transparent materials and the role of incubation." Appl. Phys. A 69 [Suppl.], S373-S376 (1999).
[CrossRef]

1997

1995

M. Chalfie, "The differentiation and function of the touch receptor neurons of Caenorhabditis elegans" Prog. Brain. Res. 105, 179-82 (1995).
[CrossRef] [PubMed]

1991

M. Driscoll and M. Chalfie, "The mec-4 gene is a member of a family of Caenorhabditis elegans genes that can mutate to induce neuronal degeneration." Nature 349, 588-593 (1991).
[CrossRef] [PubMed]

1988

1986

J.G. White, F. Southgate, J.N. Thomson and S. Brenner, "The structure of the nervous system of the nematode Caenorhabditis elegans." Phil. Trans. Royal Soc. London Series B. Bio.Scien. 314, 1-340 (1986).
[CrossRef]

1973

S. Brenner, "The genetics of behaviour," Brit. Med. Bull. 29, 269-271 (1973).
[PubMed]

1850

A. Waller, "Experiments on the section of glossopharyngeal and hypoglossal nerves of the frog and observations of the alternatives produced thereby in the structure of their primitive fibers," Philos Trans R Soc Lond Biol. 140, 423 (1850).
[CrossRef]

Adalbert, R.

B. Beirowski,R. Adalbert, D. Wagner, D.S. Grumme, K. Addicks, R.R. Ribchester and M.P. Coleman, "The progressive nature of Wallerian degeneration in wild-type and slow Wallerian degeneration (WldS) nerves," BMC Neurosci. 6 (6), (2005).
[CrossRef] [PubMed]

Addicks, K.

B. Beirowski,R. Adalbert, D. Wagner, D.S. Grumme, K. Addicks, R.R. Ribchester and M.P. Coleman, "The progressive nature of Wallerian degeneration in wild-type and slow Wallerian degeneration (WldS) nerves," BMC Neurosci. 6 (6), (2005).
[CrossRef] [PubMed]

Arakawa, N.

Ashkenasi, D.

A. Rosenfeld, M. Lorenz, R. Stoian and D. Ashkenasi, "Ultrashort-laser-pulse damage threshold of transparent materials and the role of incubation." Appl. Phys. A 69 [Suppl.], S373-S376 (1999).
[CrossRef]

Banks, P.S.

M.D. Perry, D.C. Stuart, P.S. Banks, M.D. Feit, V. Yanovsky and A.M. Rubenchik, "Ultrashort-pulse laser machining of dielectric materials," J. Appl. Phys. 85, 6803-6810 (1999).
[CrossRef]

Becker, M.F.

Beirowski, B.

B. Beirowski,R. Adalbert, D. Wagner, D.S. Grumme, K. Addicks, R.R. Ribchester and M.P. Coleman, "The progressive nature of Wallerian degeneration in wild-type and slow Wallerian degeneration (WldS) nerves," BMC Neurosci. 6 (6), (2005).
[CrossRef] [PubMed]

Ben-Yakar, A.

F. Yanik, H. Cinar, N. Cinar, A. Chisholm, Y. Jin and A. Ben-Yakar, "Nerve regeneration in Caenorhabditis elegans after femtosecond laser axotomy," IEEE J. Sel. Top. Quantum Electron. 12 (6) (2006).

M.F. Yanik, H. Cinar, H.N. Cinar, A.D. Chisholm, Y. Jin and A. Ben-Yakar, "Functional regeneration after laser axotomy," Nature 432, 882 (2004).
[CrossRef]

Bhatt, D.H.

D.H. Bhatt, S.J. Otto, B. Depoister and J.R. Fetcho, "Cyclic AMP-induced repair of zebrafish spinal circuits," Science 305, 254-258 (2004).
[CrossRef] [PubMed]

Brenner, S.

J.G. White, F. Southgate, J.N. Thomson and S. Brenner, "The structure of the nervous system of the nematode Caenorhabditis elegans." Phil. Trans. Royal Soc. London Series B. Bio.Scien. 314, 1-340 (1986).
[CrossRef]

S. Brenner, "The genetics of behaviour," Brit. Med. Bull. 29, 269-271 (1973).
[PubMed]

Chalfie, M.

M. Chalfie, "The differentiation and function of the touch receptor neurons of Caenorhabditis elegans" Prog. Brain. Res. 105, 179-82 (1995).
[CrossRef] [PubMed]

M. Driscoll and M. Chalfie, "The mec-4 gene is a member of a family of Caenorhabditis elegans genes that can mutate to induce neuronal degeneration." Nature 349, 588-593 (1991).
[CrossRef] [PubMed]

Cheng, H.J.

X. Huang, H.J. Cheng, M. Tessier-Lavigne, Y. Jin, "MAX-1, a novel PH/MyTH4/FERM domain cytoplasmic protein implicated in netrin-mediated axon repulsion" Neuron 34, 563-576 (2002).
[CrossRef] [PubMed]

Chisholm, A.

F. Yanik, H. Cinar, N. Cinar, A. Chisholm, Y. Jin and A. Ben-Yakar, "Nerve regeneration in Caenorhabditis elegans after femtosecond laser axotomy," IEEE J. Sel. Top. Quantum Electron. 12 (6) (2006).

Chisholm, A.D.

M.F. Yanik, H. Cinar, H.N. Cinar, A.D. Chisholm, Y. Jin and A. Ben-Yakar, "Functional regeneration after laser axotomy," Nature 432, 882 (2004).
[CrossRef]

Chung, S.H.

S.H. Chung, D.A. Clark, C.V. Gabel, E. Mazur and A.D.T. Samuel, "The role of the AFD neuron in C. elegans thermotaxis analyzed using femtosecond laser ablation," BMC Neuro. 7:30 (2006).

Cinar, H.

F. Yanik, H. Cinar, N. Cinar, A. Chisholm, Y. Jin and A. Ben-Yakar, "Nerve regeneration in Caenorhabditis elegans after femtosecond laser axotomy," IEEE J. Sel. Top. Quantum Electron. 12 (6) (2006).

M.F. Yanik, H. Cinar, H.N. Cinar, A.D. Chisholm, Y. Jin and A. Ben-Yakar, "Functional regeneration after laser axotomy," Nature 432, 882 (2004).
[CrossRef]

Cinar, H.N.

M.F. Yanik, H. Cinar, H.N. Cinar, A.D. Chisholm, Y. Jin and A. Ben-Yakar, "Functional regeneration after laser axotomy," Nature 432, 882 (2004).
[CrossRef]

Cinar, N.

F. Yanik, H. Cinar, N. Cinar, A. Chisholm, Y. Jin and A. Ben-Yakar, "Nerve regeneration in Caenorhabditis elegans after femtosecond laser axotomy," IEEE J. Sel. Top. Quantum Electron. 12 (6) (2006).

Clark, D.A.

S.H. Chung, D.A. Clark, C.V. Gabel, E. Mazur and A.D.T. Samuel, "The role of the AFD neuron in C. elegans thermotaxis analyzed using femtosecond laser ablation," BMC Neuro. 7:30 (2006).

Coleman, M.P.

B. Beirowski,R. Adalbert, D. Wagner, D.S. Grumme, K. Addicks, R.R. Ribchester and M.P. Coleman, "The progressive nature of Wallerian degeneration in wild-type and slow Wallerian degeneration (WldS) nerves," BMC Neurosci. 6 (6), (2005).
[CrossRef] [PubMed]

Datta, D.

N. Shen, D. Datta, C.B. Schaffer, P. LeDuc, D.E. Ingber and E. Mazur, "Ablation of cytoskeletal filaments and mitochondria in cells using a femtosecond laser nanocissor," Mech. Chem. Biosyst. 2, 17 (2005).

Depoister, B.

D.H. Bhatt, S.J. Otto, B. Depoister and J.R. Fetcho, "Cyclic AMP-induced repair of zebrafish spinal circuits," Science 305, 254-258 (2004).
[CrossRef] [PubMed]

Driscoll, M.

M. Driscoll and M. Chalfie, "The mec-4 gene is a member of a family of Caenorhabditis elegans genes that can mutate to induce neuronal degeneration." Nature 349, 588-593 (1991).
[CrossRef] [PubMed]

Feit, M.D.

M.D. Perry, D.C. Stuart, P.S. Banks, M.D. Feit, V. Yanovsky and A.M. Rubenchik, "Ultrashort-pulse laser machining of dielectric materials," J. Appl. Phys. 85, 6803-6810 (1999).
[CrossRef]

Fetcho, J.R.

D.H. Bhatt, S.J. Otto, B. Depoister and J.R. Fetcho, "Cyclic AMP-induced repair of zebrafish spinal circuits," Science 305, 254-258 (2004).
[CrossRef] [PubMed]

Fritzsche, W.

Gabel, C.V.

S.H. Chung, D.A. Clark, C.V. Gabel, E. Mazur and A.D.T. Samuel, "The role of the AFD neuron in C. elegans thermotaxis analyzed using femtosecond laser ablation," BMC Neuro. 7:30 (2006).

Gage, F.H.

P.J. Horner and F.H. Gage, "Regenerating the damaged central nervous system," Nature 407, 963-970 (2000).
[CrossRef] [PubMed]

Grumme, D.S.

B. Beirowski,R. Adalbert, D. Wagner, D.S. Grumme, K. Addicks, R.R. Ribchester and M.P. Coleman, "The progressive nature of Wallerian degeneration in wild-type and slow Wallerian degeneration (WldS) nerves," BMC Neurosci. 6 (6), (2005).
[CrossRef] [PubMed]

Guerra, A.

V. Venugopalan, A. GuerraIII, K. Hahen and A. Vogel, "Role of laser-induced plasma formation in pulse cellular microsurgery and micromanipulation," Phys. Rev. Lett. 88, 078103 (2002).
[CrossRef] [PubMed]

Guild, J.B.

Hahen, K.

V. Venugopalan, A. GuerraIII, K. Hahen and A. Vogel, "Role of laser-induced plasma formation in pulse cellular microsurgery and micromanipulation," Phys. Rev. Lett. 88, 078103 (2002).
[CrossRef] [PubMed]

Heisterkamp, A.

Horner, P.J.

P.J. Horner and F.H. Gage, "Regenerating the damaged central nervous system," Nature 407, 963-970 (2000).
[CrossRef] [PubMed]

Huang, X.

X. Huang, H.J. Cheng, M. Tessier-Lavigne, Y. Jin, "MAX-1, a novel PH/MyTH4/FERM domain cytoplasmic protein implicated in netrin-mediated axon repulsion" Neuron 34, 563-576 (2002).
[CrossRef] [PubMed]

Hüttman, G.

A. Vogel, J. Noack, G. Hüttman and G. Paltauf, "Mechanisms of femtosecond laser nanosurgery of cells and tissues," App. Phys. B, 10.1007 (2005).
[CrossRef]

Ingber, D. E.

Ingber, D.E.

N. Shen, D. Datta, C.B. Schaffer, P. LeDuc, D.E. Ingber and E. Mazur, "Ablation of cytoskeletal filaments and mitochondria in cells using a femtosecond laser nanocissor," Mech. Chem. Biosyst. 2, 17 (2005).

Jin, Y.

F. Yanik, H. Cinar, N. Cinar, A. Chisholm, Y. Jin and A. Ben-Yakar, "Nerve regeneration in Caenorhabditis elegans after femtosecond laser axotomy," IEEE J. Sel. Top. Quantum Electron. 12 (6) (2006).

M.F. Yanik, H. Cinar, H.N. Cinar, A.D. Chisholm, Y. Jin and A. Ben-Yakar, "Functional regeneration after laser axotomy," Nature 432, 882 (2004).
[CrossRef]

X. Huang, H.J. Cheng, M. Tessier-Lavigne, Y. Jin, "MAX-1, a novel PH/MyTH4/FERM domain cytoplasmic protein implicated in netrin-mediated axon repulsion" Neuron 34, 563-576 (2002).
[CrossRef] [PubMed]

Kerschensteiner, M.

M. Kerschensteiner, M.E. Schwab, J.W. Lichtman and T. Misgeld, "In vivo imaging of axonal degereneration and regeneration in the injured spinal cord," Nature Med. 11(5), 572-577 (2005).

König, K.

Kumar, S.

LeDuc, P.

N. Shen, D. Datta, C.B. Schaffer, P. LeDuc, D.E. Ingber and E. Mazur, "Ablation of cytoskeletal filaments and mitochondria in cells using a femtosecond laser nanocissor," Mech. Chem. Biosyst. 2, 17 (2005).

Lee, Y.

Lichtman, J.W.

M. Kerschensteiner, M.E. Schwab, J.W. Lichtman and T. Misgeld, "In vivo imaging of axonal degereneration and regeneration in the injured spinal cord," Nature Med. 11(5), 572-577 (2005).

Lorenz, M.

A. Rosenfeld, M. Lorenz, R. Stoian and D. Ashkenasi, "Ultrashort-laser-pulse damage threshold of transparent materials and the role of incubation." Appl. Phys. A 69 [Suppl.], S373-S376 (1999).
[CrossRef]

Mazur, E.

S.H. Chung, D.A. Clark, C.V. Gabel, E. Mazur and A.D.T. Samuel, "The role of the AFD neuron in C. elegans thermotaxis analyzed using femtosecond laser ablation," BMC Neuro. 7:30 (2006).

N. Shen, D. Datta, C.B. Schaffer, P. LeDuc, D.E. Ingber and E. Mazur, "Ablation of cytoskeletal filaments and mitochondria in cells using a femtosecond laser nanocissor," Mech. Chem. Biosyst. 2, 17 (2005).

A. Heisterkamp, I. Zaharieva Maxwell, E. Mazur, J. M. Underwood, J. A. Nickerson, S. Kumar and D. E. Ingber, "Pulse energy dependence of subcellular dissection by femtosecond laser pulses," Opt. Express 13, 3690-3696 (2005).
[CrossRef] [PubMed]

Misgeld, T.

M. Kerschensteiner, M.E. Schwab, J.W. Lichtman and T. Misgeld, "In vivo imaging of axonal degereneration and regeneration in the injured spinal cord," Nature Med. 11(5), 572-577 (2005).

Nickerson, J. A.

Noack, J.

A. Vogel, J. Noack, G. Hüttman and G. Paltauf, "Mechanisms of femtosecond laser nanosurgery of cells and tissues," App. Phys. B, 10.1007 (2005).
[CrossRef]

Otto, S.J.

D.H. Bhatt, S.J. Otto, B. Depoister and J.R. Fetcho, "Cyclic AMP-induced repair of zebrafish spinal circuits," Science 305, 254-258 (2004).
[CrossRef] [PubMed]

Paltauf, G.

A. Vogel, J. Noack, G. Hüttman and G. Paltauf, "Mechanisms of femtosecond laser nanosurgery of cells and tissues," App. Phys. B, 10.1007 (2005).
[CrossRef]

Perry, M.D.

M.D. Perry, D.C. Stuart, P.S. Banks, M.D. Feit, V. Yanovsky and A.M. Rubenchik, "Ultrashort-pulse laser machining of dielectric materials," J. Appl. Phys. 85, 6803-6810 (1999).
[CrossRef]

Ribchester, R.R.

B. Beirowski,R. Adalbert, D. Wagner, D.S. Grumme, K. Addicks, R.R. Ribchester and M.P. Coleman, "The progressive nature of Wallerian degeneration in wild-type and slow Wallerian degeneration (WldS) nerves," BMC Neurosci. 6 (6), (2005).
[CrossRef] [PubMed]

Riemann, W.

Rosenfeld, A.

A. Rosenfeld, M. Lorenz, R. Stoian and D. Ashkenasi, "Ultrashort-laser-pulse damage threshold of transparent materials and the role of incubation." Appl. Phys. A 69 [Suppl.], S373-S376 (1999).
[CrossRef]

Rubenchik, A.M.

M.D. Perry, D.C. Stuart, P.S. Banks, M.D. Feit, V. Yanovsky and A.M. Rubenchik, "Ultrashort-pulse laser machining of dielectric materials," J. Appl. Phys. 85, 6803-6810 (1999).
[CrossRef]

Samuel, A.D.T.

S.H. Chung, D.A. Clark, C.V. Gabel, E. Mazur and A.D.T. Samuel, "The role of the AFD neuron in C. elegans thermotaxis analyzed using femtosecond laser ablation," BMC Neuro. 7:30 (2006).

Schaffer, C.B.

N. Shen, D. Datta, C.B. Schaffer, P. LeDuc, D.E. Ingber and E. Mazur, "Ablation of cytoskeletal filaments and mitochondria in cells using a femtosecond laser nanocissor," Mech. Chem. Biosyst. 2, 17 (2005).

Schwab, M.E.

M. Kerschensteiner, M.E. Schwab, J.W. Lichtman and T. Misgeld, "In vivo imaging of axonal degereneration and regeneration in the injured spinal cord," Nature Med. 11(5), 572-577 (2005).

Shen, N.

N. Shen, D. Datta, C.B. Schaffer, P. LeDuc, D.E. Ingber and E. Mazur, "Ablation of cytoskeletal filaments and mitochondria in cells using a femtosecond laser nanocissor," Mech. Chem. Biosyst. 2, 17 (2005).

Southgate, F.

J.G. White, F. Southgate, J.N. Thomson and S. Brenner, "The structure of the nervous system of the nematode Caenorhabditis elegans." Phil. Trans. Royal Soc. London Series B. Bio.Scien. 314, 1-340 (1986).
[CrossRef]

Stoian, R.

A. Rosenfeld, M. Lorenz, R. Stoian and D. Ashkenasi, "Ultrashort-laser-pulse damage threshold of transparent materials and the role of incubation." Appl. Phys. A 69 [Suppl.], S373-S376 (1999).
[CrossRef]

Stuart, D.C.

M.D. Perry, D.C. Stuart, P.S. Banks, M.D. Feit, V. Yanovsky and A.M. Rubenchik, "Ultrashort-pulse laser machining of dielectric materials," J. Appl. Phys. 85, 6803-6810 (1999).
[CrossRef]

Tessier-Lavigne, M.

X. Huang, H.J. Cheng, M. Tessier-Lavigne, Y. Jin, "MAX-1, a novel PH/MyTH4/FERM domain cytoplasmic protein implicated in netrin-mediated axon repulsion" Neuron 34, 563-576 (2002).
[CrossRef] [PubMed]

Thomson, J.N.

J.G. White, F. Southgate, J.N. Thomson and S. Brenner, "The structure of the nervous system of the nematode Caenorhabditis elegans." Phil. Trans. Royal Soc. London Series B. Bio.Scien. 314, 1-340 (1986).
[CrossRef]

Tirlapur, U.K.

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

Underwood, J. M.

Urey, H.

H. Urey, "Spot size, depth of focus and diffraction ring intensity formulas for truncated Gaussian beams." App. Phys. 43 (3), 620-625 (2004).

Venugopalan, V.

V. Venugopalan, A. GuerraIII, K. Hahen and A. Vogel, "Role of laser-induced plasma formation in pulse cellular microsurgery and micromanipulation," Phys. Rev. Lett. 88, 078103 (2002).
[CrossRef] [PubMed]

Vogel, A.

A. Vogel, J. Noack, G. Hüttman and G. Paltauf, "Mechanisms of femtosecond laser nanosurgery of cells and tissues," App. Phys. B, 10.1007 (2005).
[CrossRef]

V. Venugopalan, A. GuerraIII, K. Hahen and A. Vogel, "Role of laser-induced plasma formation in pulse cellular microsurgery and micromanipulation," Phys. Rev. Lett. 88, 078103 (2002).
[CrossRef] [PubMed]

Wagner, D.

B. Beirowski,R. Adalbert, D. Wagner, D.S. Grumme, K. Addicks, R.R. Ribchester and M.P. Coleman, "The progressive nature of Wallerian degeneration in wild-type and slow Wallerian degeneration (WldS) nerves," BMC Neurosci. 6 (6), (2005).
[CrossRef] [PubMed]

Waller, A.

A. Waller, "Experiments on the section of glossopharyngeal and hypoglossal nerves of the frog and observations of the alternatives produced thereby in the structure of their primitive fibers," Philos Trans R Soc Lond Biol. 140, 423 (1850).
[CrossRef]

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Watanabe, W.

Webb, W.W.

White, J.G.

J.G. White, F. Southgate, J.N. Thomson and S. Brenner, "The structure of the nervous system of the nematode Caenorhabditis elegans." Phil. Trans. Royal Soc. London Series B. Bio.Scien. 314, 1-340 (1986).
[CrossRef]

Xu, C.

Yanik, F.

F. Yanik, H. Cinar, N. Cinar, A. Chisholm, Y. Jin and A. Ben-Yakar, "Nerve regeneration in Caenorhabditis elegans after femtosecond laser axotomy," IEEE J. Sel. Top. Quantum Electron. 12 (6) (2006).

Yanik, M.F.

M.F. Yanik, H. Cinar, H.N. Cinar, A.D. Chisholm, Y. Jin and A. Ben-Yakar, "Functional regeneration after laser axotomy," Nature 432, 882 (2004).
[CrossRef]

Yanovsky, V.

M.D. Perry, D.C. Stuart, P.S. Banks, M.D. Feit, V. Yanovsky and A.M. Rubenchik, "Ultrashort-pulse laser machining of dielectric materials," J. Appl. Phys. 85, 6803-6810 (1999).
[CrossRef]

Zaharieva Maxwell, I.

App. Phys.

H. Urey, "Spot size, depth of focus and diffraction ring intensity formulas for truncated Gaussian beams." App. Phys. 43 (3), 620-625 (2004).

App. Phys. B

A. Vogel, J. Noack, G. Hüttman and G. Paltauf, "Mechanisms of femtosecond laser nanosurgery of cells and tissues," App. Phys. B, 10.1007 (2005).
[CrossRef]

Appl. Opt.

Appl. Phys. A

A. Rosenfeld, M. Lorenz, R. Stoian and D. Ashkenasi, "Ultrashort-laser-pulse damage threshold of transparent materials and the role of incubation." Appl. Phys. A 69 [Suppl.], S373-S376 (1999).
[CrossRef]

BMC Neuro.

S.H. Chung, D.A. Clark, C.V. Gabel, E. Mazur and A.D.T. Samuel, "The role of the AFD neuron in C. elegans thermotaxis analyzed using femtosecond laser ablation," BMC Neuro. 7:30 (2006).

BMC Neurosci.

B. Beirowski,R. Adalbert, D. Wagner, D.S. Grumme, K. Addicks, R.R. Ribchester and M.P. Coleman, "The progressive nature of Wallerian degeneration in wild-type and slow Wallerian degeneration (WldS) nerves," BMC Neurosci. 6 (6), (2005).
[CrossRef] [PubMed]

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S. Brenner, "The genetics of behaviour," Brit. Med. Bull. 29, 269-271 (1973).
[PubMed]

IEEE J. Sel. Top. Quantum Electron.

F. Yanik, H. Cinar, N. Cinar, A. Chisholm, Y. Jin and A. Ben-Yakar, "Nerve regeneration in Caenorhabditis elegans after femtosecond laser axotomy," IEEE J. Sel. Top. Quantum Electron. 12 (6) (2006).

J. Appl. Phys.

M.D. Perry, D.C. Stuart, P.S. Banks, M.D. Feit, V. Yanovsky and A.M. Rubenchik, "Ultrashort-pulse laser machining of dielectric materials," J. Appl. Phys. 85, 6803-6810 (1999).
[CrossRef]

J. Opt. Soc. Am. B

Mech. Chem. Biosyst.

N. Shen, D. Datta, C.B. Schaffer, P. LeDuc, D.E. Ingber and E. Mazur, "Ablation of cytoskeletal filaments and mitochondria in cells using a femtosecond laser nanocissor," Mech. Chem. Biosyst. 2, 17 (2005).

Nature

M.F. Yanik, H. Cinar, H.N. Cinar, A.D. Chisholm, Y. Jin and A. Ben-Yakar, "Functional regeneration after laser axotomy," Nature 432, 882 (2004).
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[CrossRef] [PubMed]

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

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[CrossRef] [PubMed]

Nature Med.

M. Kerschensteiner, M.E. Schwab, J.W. Lichtman and T. Misgeld, "In vivo imaging of axonal degereneration and regeneration in the injured spinal cord," Nature Med. 11(5), 572-577 (2005).

Neuron

X. Huang, H.J. Cheng, M. Tessier-Lavigne, Y. Jin, "MAX-1, a novel PH/MyTH4/FERM domain cytoplasmic protein implicated in netrin-mediated axon repulsion" Neuron 34, 563-576 (2002).
[CrossRef] [PubMed]

Opt. Express

Opt. Lett.

Philos Trans R Soc Lond Biol.

A. Waller, "Experiments on the section of glossopharyngeal and hypoglossal nerves of the frog and observations of the alternatives produced thereby in the structure of their primitive fibers," Philos Trans R Soc Lond Biol. 140, 423 (1850).
[CrossRef]

Phys. Rev. Lett.

V. Venugopalan, A. GuerraIII, K. Hahen and A. Vogel, "Role of laser-induced plasma formation in pulse cellular microsurgery and micromanipulation," Phys. Rev. Lett. 88, 078103 (2002).
[CrossRef] [PubMed]

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M. Chalfie, "The differentiation and function of the touch receptor neurons of Caenorhabditis elegans" Prog. Brain. Res. 105, 179-82 (1995).
[CrossRef] [PubMed]

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J.G. White, F. Southgate, J.N. Thomson and S. Brenner, "The structure of the nervous system of the nematode Caenorhabditis elegans." Phil. Trans. Royal Soc. London Series B. Bio.Scien. 314, 1-340 (1986).
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Science

D.H. Bhatt, S.J. Otto, B. Depoister and J.R. Fetcho, "Cyclic AMP-induced repair of zebrafish spinal circuits," Science 305, 254-258 (2004).
[CrossRef] [PubMed]

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W.B. Wood, S. Brenner, R.K. Herman, S.W. Emmons, J. White, J. Sulston, H.R. Horvitz, J. Kimble, S. Ward, J. Hodgkin, R.H. Waterston, M. Chalfie and D.L. Riddle, The nematode Caenorhabditis elegans, (Cold Spring Harbor, 1988).

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Figures (10)

Fig. 1.
Fig. 1.

(a) Sketch of all GABA-ergic neurons in C. elegans (head to the right). The 13 VDs and the 6 DDs are the motor neurons. The insert shows myosin filaments underlying an axon [17]. (b) Sketch of soft touch neurons in C. elegans (head to the left) [18]. ALM and PLM come in pairs; only one of each is accessible during surgery.

Fig. 2.
Fig. 2.

Setup where nanosurgery is incorporated in a fluorescence microscope.

Fig. 3.
Fig. 3.

Fluorescence images of an axon of a GFP-labeled PLML neuron (a) before axotomy, (b) right after the axotomy at mid-body (100 pulses of 1.9 J/cm2), and (c) 12 hours after the axotomy showing regrowth of the severed axon and reconnection to its distal end.

Fig. 4.
Fig. 4.

Images of regrown ALML axon 72 hours after axotomy on a youg adult worm (300 pulses of 1.9 J/cm2) at three different focal depths. The first two images show the connection to the degenerated distal end and the third image shows how the process continues to regrow most probably in search of a healthier connection. (The distal and proximal ends are respectively on the upper and lower parts of the pictures).

Fig. 5.
Fig. 5.

Fluorescence images of photobleaching at 1.3 J/cm2, 25 pulses on CZ5062 , (a) before laser exposure, (b) right after exposure, and (c) 2 minutes after exposure. The irradiated spot loses its signal due to photobleaching but remains intact. The axon recovers its luminescence within 2 minutes by the GFP diffusion within the cytoplasm.

Fig. 6.
Fig. 6.

Fluorescence images of the extent of damage induced by fs-laser ablation using 800 pulses at 1.7 J/cm2. (a) before ablation, (b) right after ablation with GFP spilling in the created cavity, and (c) 3 minutes after ablation. The dark region in muscle fibers shows the extent of damage to surrounding tissue. The diameter of this area is approximately equal to the distance between the ends of the cut axon.

Fig. 7.
Fig. 7.

Ablation thresholds measured as a function of number of pulses. Solid line shows the logarithmic fit following Eq. (2). The corresponding fluences (energy per area) and irradiances are included in the right side axis assuming a theoretical spot size of 620 nm and a pulse duration of 430 fs. The error bars indicate the variance of the threshold measurements on 10 axons for different pulse trains.

Fig. 8.
Fig. 8.

Extent of photo-damage induced by fs-laser ablation of axons in C. elegans as a function of pulse energy and total number of pulses.

Fig. 9.
Fig. 9.

Fluorescence images of the extent of damage, (a-d) at 8 nJ for 100, 200, 400 and 800 pulses, (e-h) for 400 pulses at 6, 8, 10 and 12 nJ.

Fig. 10.
Fig. 10.

Axonal recovery probabilities of touch neurons. Red bars on the left are survival rates after anesthesia and surgery. Green bars on the right are axonal recovery rates.

Tables (2)

Tables Icon

Table 1. Damage mechanisms in fs-laser nanosurgery at kHz repetition rate [8]. The irradiance thresholds and the free electron densities were estimated by Vogel et al. for 100 fs, 800 nm laser pulses focused with 1.3 NA lens [8].

Tables Icon

Table 2. Statistics of axonal recovery of touch neurons and survival rate of worms after laser axotomy performed using different pulse energies and total number of pulses. Relative rates refer to the number of worms fulfilling the requirement of the previous column.

Equations (2)

Equations on this page are rendered with MathJax. Learn more.

F N = F 1 + ( F F 1 ) N k ,
E N = E 1 + ( E E 1 ) N k ,

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