Abstract

Nanosecond laser pulses (λ=355 nm) were used to cut mechanosensory neurons in Caenorhabditis elegans and motorneurons in Drosophila melanogaster larvae. A pulse energy range of 0.8–1.2 µJ and <20 pulses in single shot mode were sufficient to generate axonal cuts. Viability post-surgery was >95% for C. elegans and 60% for Drosophila. Cavitation bubble dynamics generated due to laser-induced plasma formation were observed in vivo by time-resolved imaging in both organisms. Bubble oscillations were severely damped in vivo and cavitation dynamics were complete within 100 ns in C. elegans and 800 ns in Drosophila. We report the use of this system to study axonal transport for the first time and discuss advantages of nanosecond lasers compared to femtosecond sources for such procedures.

© 2008 Optical Society of America

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  1. 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]
  2. S. H. Chung, D. A. Clark, C. V. Gabel, E. Mazur, and A. D. Samuel,"The role of the AFD neuron in C. elegans thermotaxis analyzed using femtosecond laser ablation," BMC Neuroscience 7, DOI:10.1186/1471-2202-7-30 (2006).
    [CrossRef] [PubMed]
  3. Z. Wu, A. Ghosh-Roy, M. F. Yanik, J. Z. Zhang, Y. Jin, and A. D. Chisholm, "Caenorhabditis elegans neuronal regeneration is influenced by life stage, ephrin signaling and synaptic branching," PNAS 104,15132-15137 (2007).
    [CrossRef] [PubMed]
  4. P. Weiss and H. B. Hiscoe,"Experiments on the mechanism of nerve growth," J. Exp. Zool. 107315-395 (1948).
    [CrossRef] [PubMed]
  5. R. V. Barkus, O. Klyachko, D. Horiuchi B. J. Dickson and W. M. Saxton, "Identification of an Axonal Kinesin-3 Motor for Fast Anterograde Vesicle Transport that Facilitates Retrograde Transport of Neuropeptides," Mol. Biol. Cell 19,274-283 (2008).
    [CrossRef]
  6. H. Erez, G. Malkinson, M. Prager-Khoutorsky, C. I. De Zeeuw, C. C. Hoogenraad, and M. E. Spira, "Formation of microtubule-based traps controls the sorting and concentration of vesicles to restricted sites of regenerating neurons after axotomy," J. Cell Biol. 176,497-507 (2007).
    [CrossRef] [PubMed]
  7. M. S. Hutson and X. Ma, "Plasma and cavitation dynamics during pulsed laser microsurgery in vivo," Phys. Rev. Lett. 99,158104 (2007).
    [CrossRef] [PubMed]
  8. T. R. Mahoney, Q. Liu, T. Itoh, S. Luo, G. Hadwiger, R. Vincent, Z-W. Wang, M. Fukuda, and M. L. Nonet, "Regulation of synaptic transmission by RAB-3 and RAB-27 in Caenorhabditis elegans," Mol. Biol. Cell 17,2617-2625 (2006).
    [CrossRef] [PubMed]
  9. M. L. Nonet, "Visualization of synaptic specializations in live C. elegans with synaptic vesicle protein-GFP fusions," J. Neurosci. Methods 89,33-40 (1999).
    [CrossRef] [PubMed]
  10. T. Lee and L. Luo, "Mosaic analysis with a repressible cell marker for studies of gene function in neuronal morphogenesis," Neuron 22,451-61 (1999).
    [CrossRef] [PubMed]
  11. A. Mahr and H. Aberle, "The expression pattern of the Drosophila vesicular glutamate transporter: A marker protein for motoneurons and glutamatergic centers in the brain, Gene Expr. Patterns 6,299-309 (2006).
    [CrossRef]
  12. A. V. Cherian and K. R. Rau, "Pulsed laser-induced damage in rat corneas: time-resolved imaging of physical effects and acute biological response," J. Biomed. Opt. 13,024009 (2008).
    [CrossRef] [PubMed]
  13. M. F. Yanik, H. Cinar, H. N. Cinar, A. Gibby, A. D. Chisholm, Y. Jin, and A. Ben-Yakar, "Nerve regeneration in Caenorhabditis elegans after femtosecond laser axotomy," IEEE J. Quantum Electron. 12,1283-1291 (2006).
    [CrossRef]
  14. V. Venugopalan, A. GuerraIII, K. Nahen, and A. Vogel, "Role of Laser-Induced Plasma Formation in Pulsed Cellular Microsurgery and Micromanipulation," Phys. Rev. Lett. 88,078103 (2002).
    [CrossRef] [PubMed]
  15. U. K. Tirlapur and K. Konig, "Targeted transfection by femtosecond laser," Nature 418,290-291 (2002).
    [CrossRef] [PubMed]
  16. W. Watanabe and N. Arakawa, "Femtosecond laser disruption of subcellular organelles in a living cell," Opt. Express 12,4203-4213 (2004).
    [CrossRef] [PubMed]
  17. 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]
  18. V. Magidson, J. Loncarek, P. Hergert, C. L. Reider, and A. Khodjakov,"Laser Microsurgery in the GFP Era: A Cell Biologist???s Perspective," Methods Cell Biol. 82,239-266, (2007).
    [PubMed]
  19. W. Grill, P. Gonczy, E. H. K. Stelzer, and A. A. Hyman "Polarity controls forces governing asymmetric spindle positioning in the Caenorhabditis elegans embryo," Nature 409,630-633 (2001).
    [CrossRef] [PubMed]
  20. M. S. Hutson, Y. Tokutake, M.-S. Chang, J.W. Bloor, S. Venakides, D. P. Kiehart, and G. S. Edwards "Forces for morphogenesis investigated with laser-microsurgery and quantitative modeling," Science 300,145-149 (2003).
    [CrossRef] [PubMed]
  21. F. Bourgeois and A. Ben-Yakar, "Femtosecond laser nanoaxotomy properties and their effect on axonal recovery in C. elegans," Opt. Express 15,8521-8531 (2007).
    [CrossRef] [PubMed]
  22. J. J. Fernandes and H. Keshishian "Nerve-muscle interactions during flight muscle development in Drosophila," Development,  125,1769-1779 (1998).
    [PubMed]
  23. K. R. Rau, P. A. Quinto-Su, A. Hellman, and V. Venugopalan, "Pulsed laser microbeam-induced cell lysis: timeresolved imaging and analysis of hydrodynamic effects," Biophys. J. 91,317-329 (2006).
    [CrossRef] [PubMed]
  24. A. Vogel, M. R. C. Capon, M. N. Asiyo-Vogel, and R. Birngruber, "Intraocular photodisruption with picosecond and nanosecond laser pulses: Tissue effects in cornea, lens, and retina," Invest. Ophthalmol. Visual Sci. 35,3032-3044 (1994).
  25. A. Vogel, N. Linz, S. Freidank, and G. Paltauf, "Femtosecond-Laser-Induced Nanocavitation in Water: Implications for Optical Breakdown Threshold and Cell Surgery," Phys. Rev. Lett. 100,038102 (2008).
    [CrossRef] [PubMed]

2008

R. V. Barkus, O. Klyachko, D. Horiuchi B. J. Dickson and W. M. Saxton, "Identification of an Axonal Kinesin-3 Motor for Fast Anterograde Vesicle Transport that Facilitates Retrograde Transport of Neuropeptides," Mol. Biol. Cell 19,274-283 (2008).
[CrossRef]

A. V. Cherian and K. R. Rau, "Pulsed laser-induced damage in rat corneas: time-resolved imaging of physical effects and acute biological response," J. Biomed. Opt. 13,024009 (2008).
[CrossRef] [PubMed]

A. Vogel, N. Linz, S. Freidank, and G. Paltauf, "Femtosecond-Laser-Induced Nanocavitation in Water: Implications for Optical Breakdown Threshold and Cell Surgery," Phys. Rev. Lett. 100,038102 (2008).
[CrossRef] [PubMed]

2007

F. Bourgeois and A. Ben-Yakar, "Femtosecond laser nanoaxotomy properties and their effect on axonal recovery in C. elegans," Opt. Express 15,8521-8531 (2007).
[CrossRef] [PubMed]

V. Magidson, J. Loncarek, P. Hergert, C. L. Reider, and A. Khodjakov,"Laser Microsurgery in the GFP Era: A Cell Biologist???s Perspective," Methods Cell Biol. 82,239-266, (2007).
[PubMed]

H. Erez, G. Malkinson, M. Prager-Khoutorsky, C. I. De Zeeuw, C. C. Hoogenraad, and M. E. Spira, "Formation of microtubule-based traps controls the sorting and concentration of vesicles to restricted sites of regenerating neurons after axotomy," J. Cell Biol. 176,497-507 (2007).
[CrossRef] [PubMed]

M. S. Hutson and X. Ma, "Plasma and cavitation dynamics during pulsed laser microsurgery in vivo," Phys. Rev. Lett. 99,158104 (2007).
[CrossRef] [PubMed]

Z. Wu, A. Ghosh-Roy, M. F. Yanik, J. Z. Zhang, Y. Jin, and A. D. Chisholm, "Caenorhabditis elegans neuronal regeneration is influenced by life stage, ephrin signaling and synaptic branching," PNAS 104,15132-15137 (2007).
[CrossRef] [PubMed]

2006

S. H. Chung, D. A. Clark, C. V. Gabel, E. Mazur, and A. D. Samuel,"The role of the AFD neuron in C. elegans thermotaxis analyzed using femtosecond laser ablation," BMC Neuroscience 7, DOI:10.1186/1471-2202-7-30 (2006).
[CrossRef] [PubMed]

T. R. Mahoney, Q. Liu, T. Itoh, S. Luo, G. Hadwiger, R. Vincent, Z-W. Wang, M. Fukuda, and M. L. Nonet, "Regulation of synaptic transmission by RAB-3 and RAB-27 in Caenorhabditis elegans," Mol. Biol. Cell 17,2617-2625 (2006).
[CrossRef] [PubMed]

M. F. Yanik, H. Cinar, H. N. Cinar, A. Gibby, A. D. Chisholm, Y. Jin, and A. Ben-Yakar, "Nerve regeneration in Caenorhabditis elegans after femtosecond laser axotomy," IEEE J. Quantum Electron. 12,1283-1291 (2006).
[CrossRef]

A. Mahr and H. Aberle, "The expression pattern of the Drosophila vesicular glutamate transporter: A marker protein for motoneurons and glutamatergic centers in the brain, Gene Expr. Patterns 6,299-309 (2006).
[CrossRef]

K. R. Rau, P. A. Quinto-Su, A. Hellman, and V. Venugopalan, "Pulsed laser microbeam-induced cell lysis: timeresolved imaging and analysis of hydrodynamic effects," Biophys. J. 91,317-329 (2006).
[CrossRef] [PubMed]

2005

2004

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

2003

M. S. Hutson, Y. Tokutake, M.-S. Chang, J.W. Bloor, S. Venakides, D. P. Kiehart, and G. S. Edwards "Forces for morphogenesis investigated with laser-microsurgery and quantitative modeling," Science 300,145-149 (2003).
[CrossRef] [PubMed]

2002

V. Venugopalan, A. GuerraIII, K. Nahen, and A. Vogel, "Role of Laser-Induced Plasma Formation in Pulsed Cellular Microsurgery and Micromanipulation," Phys. Rev. Lett. 88,078103 (2002).
[CrossRef] [PubMed]

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

2001

W. Grill, P. Gonczy, E. H. K. Stelzer, and A. A. Hyman "Polarity controls forces governing asymmetric spindle positioning in the Caenorhabditis elegans embryo," Nature 409,630-633 (2001).
[CrossRef] [PubMed]

1999

M. L. Nonet, "Visualization of synaptic specializations in live C. elegans with synaptic vesicle protein-GFP fusions," J. Neurosci. Methods 89,33-40 (1999).
[CrossRef] [PubMed]

T. Lee and L. Luo, "Mosaic analysis with a repressible cell marker for studies of gene function in neuronal morphogenesis," Neuron 22,451-61 (1999).
[CrossRef] [PubMed]

1998

J. J. Fernandes and H. Keshishian "Nerve-muscle interactions during flight muscle development in Drosophila," Development,  125,1769-1779 (1998).
[PubMed]

1994

A. Vogel, M. R. C. Capon, M. N. Asiyo-Vogel, and R. Birngruber, "Intraocular photodisruption with picosecond and nanosecond laser pulses: Tissue effects in cornea, lens, and retina," Invest. Ophthalmol. Visual Sci. 35,3032-3044 (1994).

1948

P. Weiss and H. B. Hiscoe,"Experiments on the mechanism of nerve growth," J. Exp. Zool. 107315-395 (1948).
[CrossRef] [PubMed]

Aberle, H.

A. Mahr and H. Aberle, "The expression pattern of the Drosophila vesicular glutamate transporter: A marker protein for motoneurons and glutamatergic centers in the brain, Gene Expr. Patterns 6,299-309 (2006).
[CrossRef]

Arakawa, N.

Asiyo-Vogel, M. N.

A. Vogel, M. R. C. Capon, M. N. Asiyo-Vogel, and R. Birngruber, "Intraocular photodisruption with picosecond and nanosecond laser pulses: Tissue effects in cornea, lens, and retina," Invest. Ophthalmol. Visual Sci. 35,3032-3044 (1994).

Barkus, R. V.

R. V. Barkus, O. Klyachko, D. Horiuchi B. J. Dickson and W. M. Saxton, "Identification of an Axonal Kinesin-3 Motor for Fast Anterograde Vesicle Transport that Facilitates Retrograde Transport of Neuropeptides," Mol. Biol. Cell 19,274-283 (2008).
[CrossRef]

Ben-Yakar, A.

F. Bourgeois and A. Ben-Yakar, "Femtosecond laser nanoaxotomy properties and their effect on axonal recovery in C. elegans," Opt. Express 15,8521-8531 (2007).
[CrossRef] [PubMed]

M. F. Yanik, H. Cinar, H. N. Cinar, A. Gibby, A. D. Chisholm, Y. Jin, and A. Ben-Yakar, "Nerve regeneration in Caenorhabditis elegans after femtosecond laser axotomy," IEEE J. Quantum Electron. 12,1283-1291 (2006).
[CrossRef]

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]

Birngruber, R.

A. Vogel, M. R. C. Capon, M. N. Asiyo-Vogel, and R. Birngruber, "Intraocular photodisruption with picosecond and nanosecond laser pulses: Tissue effects in cornea, lens, and retina," Invest. Ophthalmol. Visual Sci. 35,3032-3044 (1994).

Bloor, J. W.

M. S. Hutson, Y. Tokutake, M.-S. Chang, J.W. Bloor, S. Venakides, D. P. Kiehart, and G. S. Edwards "Forces for morphogenesis investigated with laser-microsurgery and quantitative modeling," Science 300,145-149 (2003).
[CrossRef] [PubMed]

Bourgeois, F.

Capon, M. R. C.

A. Vogel, M. R. C. Capon, M. N. Asiyo-Vogel, and R. Birngruber, "Intraocular photodisruption with picosecond and nanosecond laser pulses: Tissue effects in cornea, lens, and retina," Invest. Ophthalmol. Visual Sci. 35,3032-3044 (1994).

Chang, M.-S.

M. S. Hutson, Y. Tokutake, M.-S. Chang, J.W. Bloor, S. Venakides, D. P. Kiehart, and G. S. Edwards "Forces for morphogenesis investigated with laser-microsurgery and quantitative modeling," Science 300,145-149 (2003).
[CrossRef] [PubMed]

Cherian, A. V.

A. V. Cherian and K. R. Rau, "Pulsed laser-induced damage in rat corneas: time-resolved imaging of physical effects and acute biological response," J. Biomed. Opt. 13,024009 (2008).
[CrossRef] [PubMed]

Chisholm, A. D.

Z. Wu, A. Ghosh-Roy, M. F. Yanik, J. Z. Zhang, Y. Jin, and A. D. Chisholm, "Caenorhabditis elegans neuronal regeneration is influenced by life stage, ephrin signaling and synaptic branching," PNAS 104,15132-15137 (2007).
[CrossRef] [PubMed]

M. F. Yanik, H. Cinar, H. N. Cinar, A. Gibby, A. D. Chisholm, Y. Jin, and A. Ben-Yakar, "Nerve regeneration in Caenorhabditis elegans after femtosecond laser axotomy," IEEE J. Quantum Electron. 12,1283-1291 (2006).
[CrossRef]

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. Samuel,"The role of the AFD neuron in C. elegans thermotaxis analyzed using femtosecond laser ablation," BMC Neuroscience 7, DOI:10.1186/1471-2202-7-30 (2006).
[CrossRef] [PubMed]

Cinar, H.

M. F. Yanik, H. Cinar, H. N. Cinar, A. Gibby, A. D. Chisholm, Y. Jin, and A. Ben-Yakar, "Nerve regeneration in Caenorhabditis elegans after femtosecond laser axotomy," IEEE J. Quantum Electron. 12,1283-1291 (2006).
[CrossRef]

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. Gibby, A. D. Chisholm, Y. Jin, and A. Ben-Yakar, "Nerve regeneration in Caenorhabditis elegans after femtosecond laser axotomy," IEEE J. Quantum Electron. 12,1283-1291 (2006).
[CrossRef]

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]

Clark, D. A.

S. H. Chung, D. A. Clark, C. V. Gabel, E. Mazur, and A. D. Samuel,"The role of the AFD neuron in C. elegans thermotaxis analyzed using femtosecond laser ablation," BMC Neuroscience 7, DOI:10.1186/1471-2202-7-30 (2006).
[CrossRef] [PubMed]

De Zeeuw, C. I.

H. Erez, G. Malkinson, M. Prager-Khoutorsky, C. I. De Zeeuw, C. C. Hoogenraad, and M. E. Spira, "Formation of microtubule-based traps controls the sorting and concentration of vesicles to restricted sites of regenerating neurons after axotomy," J. Cell Biol. 176,497-507 (2007).
[CrossRef] [PubMed]

Edwards, G. S.

M. S. Hutson, Y. Tokutake, M.-S. Chang, J.W. Bloor, S. Venakides, D. P. Kiehart, and G. S. Edwards "Forces for morphogenesis investigated with laser-microsurgery and quantitative modeling," Science 300,145-149 (2003).
[CrossRef] [PubMed]

Erez, H.

H. Erez, G. Malkinson, M. Prager-Khoutorsky, C. I. De Zeeuw, C. C. Hoogenraad, and M. E. Spira, "Formation of microtubule-based traps controls the sorting and concentration of vesicles to restricted sites of regenerating neurons after axotomy," J. Cell Biol. 176,497-507 (2007).
[CrossRef] [PubMed]

Fernandes, J. J.

J. J. Fernandes and H. Keshishian "Nerve-muscle interactions during flight muscle development in Drosophila," Development,  125,1769-1779 (1998).
[PubMed]

Freidank, S.

A. Vogel, N. Linz, S. Freidank, and G. Paltauf, "Femtosecond-Laser-Induced Nanocavitation in Water: Implications for Optical Breakdown Threshold and Cell Surgery," Phys. Rev. Lett. 100,038102 (2008).
[CrossRef] [PubMed]

Fukuda, M.

T. R. Mahoney, Q. Liu, T. Itoh, S. Luo, G. Hadwiger, R. Vincent, Z-W. Wang, M. Fukuda, and M. L. Nonet, "Regulation of synaptic transmission by RAB-3 and RAB-27 in Caenorhabditis elegans," Mol. Biol. Cell 17,2617-2625 (2006).
[CrossRef] [PubMed]

Gabel, C. V.

S. H. Chung, D. A. Clark, C. V. Gabel, E. Mazur, and A. D. Samuel,"The role of the AFD neuron in C. elegans thermotaxis analyzed using femtosecond laser ablation," BMC Neuroscience 7, DOI:10.1186/1471-2202-7-30 (2006).
[CrossRef] [PubMed]

Ghosh-Roy, A.

Z. Wu, A. Ghosh-Roy, M. F. Yanik, J. Z. Zhang, Y. Jin, and A. D. Chisholm, "Caenorhabditis elegans neuronal regeneration is influenced by life stage, ephrin signaling and synaptic branching," PNAS 104,15132-15137 (2007).
[CrossRef] [PubMed]

Gibby, A.

M. F. Yanik, H. Cinar, H. N. Cinar, A. Gibby, A. D. Chisholm, Y. Jin, and A. Ben-Yakar, "Nerve regeneration in Caenorhabditis elegans after femtosecond laser axotomy," IEEE J. Quantum Electron. 12,1283-1291 (2006).
[CrossRef]

Gonczy, P.

W. Grill, P. Gonczy, E. H. K. Stelzer, and A. A. Hyman "Polarity controls forces governing asymmetric spindle positioning in the Caenorhabditis elegans embryo," Nature 409,630-633 (2001).
[CrossRef] [PubMed]

Grill, W.

W. Grill, P. Gonczy, E. H. K. Stelzer, and A. A. Hyman "Polarity controls forces governing asymmetric spindle positioning in the Caenorhabditis elegans embryo," Nature 409,630-633 (2001).
[CrossRef] [PubMed]

Guerra, A.

V. Venugopalan, A. GuerraIII, K. Nahen, and A. Vogel, "Role of Laser-Induced Plasma Formation in Pulsed Cellular Microsurgery and Micromanipulation," Phys. Rev. Lett. 88,078103 (2002).
[CrossRef] [PubMed]

Hadwiger, G.

T. R. Mahoney, Q. Liu, T. Itoh, S. Luo, G. Hadwiger, R. Vincent, Z-W. Wang, M. Fukuda, and M. L. Nonet, "Regulation of synaptic transmission by RAB-3 and RAB-27 in Caenorhabditis elegans," Mol. Biol. Cell 17,2617-2625 (2006).
[CrossRef] [PubMed]

Heisterkamp, A.

Hellman, A.

K. R. Rau, P. A. Quinto-Su, A. Hellman, and V. Venugopalan, "Pulsed laser microbeam-induced cell lysis: timeresolved imaging and analysis of hydrodynamic effects," Biophys. J. 91,317-329 (2006).
[CrossRef] [PubMed]

Hergert, P.

V. Magidson, J. Loncarek, P. Hergert, C. L. Reider, and A. Khodjakov,"Laser Microsurgery in the GFP Era: A Cell Biologist???s Perspective," Methods Cell Biol. 82,239-266, (2007).
[PubMed]

Hiscoe, H. B.

P. Weiss and H. B. Hiscoe,"Experiments on the mechanism of nerve growth," J. Exp. Zool. 107315-395 (1948).
[CrossRef] [PubMed]

Hoogenraad, C. C.

H. Erez, G. Malkinson, M. Prager-Khoutorsky, C. I. De Zeeuw, C. C. Hoogenraad, and M. E. Spira, "Formation of microtubule-based traps controls the sorting and concentration of vesicles to restricted sites of regenerating neurons after axotomy," J. Cell Biol. 176,497-507 (2007).
[CrossRef] [PubMed]

Hutson, M. S.

M. S. Hutson and X. Ma, "Plasma and cavitation dynamics during pulsed laser microsurgery in vivo," Phys. Rev. Lett. 99,158104 (2007).
[CrossRef] [PubMed]

M. S. Hutson, Y. Tokutake, M.-S. Chang, J.W. Bloor, S. Venakides, D. P. Kiehart, and G. S. Edwards "Forces for morphogenesis investigated with laser-microsurgery and quantitative modeling," Science 300,145-149 (2003).
[CrossRef] [PubMed]

Hyman, A. A.

W. Grill, P. Gonczy, E. H. K. Stelzer, and A. A. Hyman "Polarity controls forces governing asymmetric spindle positioning in the Caenorhabditis elegans embryo," Nature 409,630-633 (2001).
[CrossRef] [PubMed]

Ingber, D. E.

Itoh, T.

T. R. Mahoney, Q. Liu, T. Itoh, S. Luo, G. Hadwiger, R. Vincent, Z-W. Wang, M. Fukuda, and M. L. Nonet, "Regulation of synaptic transmission by RAB-3 and RAB-27 in Caenorhabditis elegans," Mol. Biol. Cell 17,2617-2625 (2006).
[CrossRef] [PubMed]

Jin, Y.

Z. Wu, A. Ghosh-Roy, M. F. Yanik, J. Z. Zhang, Y. Jin, and A. D. Chisholm, "Caenorhabditis elegans neuronal regeneration is influenced by life stage, ephrin signaling and synaptic branching," PNAS 104,15132-15137 (2007).
[CrossRef] [PubMed]

M. F. Yanik, H. Cinar, H. N. Cinar, A. Gibby, A. D. Chisholm, Y. Jin, and A. Ben-Yakar, "Nerve regeneration in Caenorhabditis elegans after femtosecond laser axotomy," IEEE J. Quantum Electron. 12,1283-1291 (2006).
[CrossRef]

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]

Keshishian, H.

J. J. Fernandes and H. Keshishian "Nerve-muscle interactions during flight muscle development in Drosophila," Development,  125,1769-1779 (1998).
[PubMed]

Khodjakov, A.

V. Magidson, J. Loncarek, P. Hergert, C. L. Reider, and A. Khodjakov,"Laser Microsurgery in the GFP Era: A Cell Biologist???s Perspective," Methods Cell Biol. 82,239-266, (2007).
[PubMed]

Kiehart, D. P.

M. S. Hutson, Y. Tokutake, M.-S. Chang, J.W. Bloor, S. Venakides, D. P. Kiehart, and G. S. Edwards "Forces for morphogenesis investigated with laser-microsurgery and quantitative modeling," Science 300,145-149 (2003).
[CrossRef] [PubMed]

Klyachko, O.

R. V. Barkus, O. Klyachko, D. Horiuchi B. J. Dickson and W. M. Saxton, "Identification of an Axonal Kinesin-3 Motor for Fast Anterograde Vesicle Transport that Facilitates Retrograde Transport of Neuropeptides," Mol. Biol. Cell 19,274-283 (2008).
[CrossRef]

Konig, K.

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

Kumar, S.

Lee, T.

T. Lee and L. Luo, "Mosaic analysis with a repressible cell marker for studies of gene function in neuronal morphogenesis," Neuron 22,451-61 (1999).
[CrossRef] [PubMed]

Linz, N.

A. Vogel, N. Linz, S. Freidank, and G. Paltauf, "Femtosecond-Laser-Induced Nanocavitation in Water: Implications for Optical Breakdown Threshold and Cell Surgery," Phys. Rev. Lett. 100,038102 (2008).
[CrossRef] [PubMed]

Liu, Q.

T. R. Mahoney, Q. Liu, T. Itoh, S. Luo, G. Hadwiger, R. Vincent, Z-W. Wang, M. Fukuda, and M. L. Nonet, "Regulation of synaptic transmission by RAB-3 and RAB-27 in Caenorhabditis elegans," Mol. Biol. Cell 17,2617-2625 (2006).
[CrossRef] [PubMed]

Loncarek, J.

V. Magidson, J. Loncarek, P. Hergert, C. L. Reider, and A. Khodjakov,"Laser Microsurgery in the GFP Era: A Cell Biologist???s Perspective," Methods Cell Biol. 82,239-266, (2007).
[PubMed]

Luo, L.

T. Lee and L. Luo, "Mosaic analysis with a repressible cell marker for studies of gene function in neuronal morphogenesis," Neuron 22,451-61 (1999).
[CrossRef] [PubMed]

Luo, S.

T. R. Mahoney, Q. Liu, T. Itoh, S. Luo, G. Hadwiger, R. Vincent, Z-W. Wang, M. Fukuda, and M. L. Nonet, "Regulation of synaptic transmission by RAB-3 and RAB-27 in Caenorhabditis elegans," Mol. Biol. Cell 17,2617-2625 (2006).
[CrossRef] [PubMed]

Ma, X.

M. S. Hutson and X. Ma, "Plasma and cavitation dynamics during pulsed laser microsurgery in vivo," Phys. Rev. Lett. 99,158104 (2007).
[CrossRef] [PubMed]

Magidson, V.

V. Magidson, J. Loncarek, P. Hergert, C. L. Reider, and A. Khodjakov,"Laser Microsurgery in the GFP Era: A Cell Biologist???s Perspective," Methods Cell Biol. 82,239-266, (2007).
[PubMed]

Mahoney, T. R.

T. R. Mahoney, Q. Liu, T. Itoh, S. Luo, G. Hadwiger, R. Vincent, Z-W. Wang, M. Fukuda, and M. L. Nonet, "Regulation of synaptic transmission by RAB-3 and RAB-27 in Caenorhabditis elegans," Mol. Biol. Cell 17,2617-2625 (2006).
[CrossRef] [PubMed]

Mahr, A.

A. Mahr and H. Aberle, "The expression pattern of the Drosophila vesicular glutamate transporter: A marker protein for motoneurons and glutamatergic centers in the brain, Gene Expr. Patterns 6,299-309 (2006).
[CrossRef]

Malkinson, G.

H. Erez, G. Malkinson, M. Prager-Khoutorsky, C. I. De Zeeuw, C. C. Hoogenraad, and M. E. Spira, "Formation of microtubule-based traps controls the sorting and concentration of vesicles to restricted sites of regenerating neurons after axotomy," J. Cell Biol. 176,497-507 (2007).
[CrossRef] [PubMed]

Mazur, E.

S. H. Chung, D. A. Clark, C. V. Gabel, E. Mazur, and A. D. Samuel,"The role of the AFD neuron in C. elegans thermotaxis analyzed using femtosecond laser ablation," BMC Neuroscience 7, DOI:10.1186/1471-2202-7-30 (2006).
[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]

Nahen, K.

V. Venugopalan, A. GuerraIII, K. Nahen, and A. Vogel, "Role of Laser-Induced Plasma Formation in Pulsed Cellular Microsurgery and Micromanipulation," Phys. Rev. Lett. 88,078103 (2002).
[CrossRef] [PubMed]

Nickerson, J. A.

Nonet, M. L.

T. R. Mahoney, Q. Liu, T. Itoh, S. Luo, G. Hadwiger, R. Vincent, Z-W. Wang, M. Fukuda, and M. L. Nonet, "Regulation of synaptic transmission by RAB-3 and RAB-27 in Caenorhabditis elegans," Mol. Biol. Cell 17,2617-2625 (2006).
[CrossRef] [PubMed]

M. L. Nonet, "Visualization of synaptic specializations in live C. elegans with synaptic vesicle protein-GFP fusions," J. Neurosci. Methods 89,33-40 (1999).
[CrossRef] [PubMed]

Paltauf, G.

A. Vogel, N. Linz, S. Freidank, and G. Paltauf, "Femtosecond-Laser-Induced Nanocavitation in Water: Implications for Optical Breakdown Threshold and Cell Surgery," Phys. Rev. Lett. 100,038102 (2008).
[CrossRef] [PubMed]

Prager-Khoutorsky, M.

H. Erez, G. Malkinson, M. Prager-Khoutorsky, C. I. De Zeeuw, C. C. Hoogenraad, and M. E. Spira, "Formation of microtubule-based traps controls the sorting and concentration of vesicles to restricted sites of regenerating neurons after axotomy," J. Cell Biol. 176,497-507 (2007).
[CrossRef] [PubMed]

Quinto-Su, P. A.

K. R. Rau, P. A. Quinto-Su, A. Hellman, and V. Venugopalan, "Pulsed laser microbeam-induced cell lysis: timeresolved imaging and analysis of hydrodynamic effects," Biophys. J. 91,317-329 (2006).
[CrossRef] [PubMed]

Rau, K. R.

A. V. Cherian and K. R. Rau, "Pulsed laser-induced damage in rat corneas: time-resolved imaging of physical effects and acute biological response," J. Biomed. Opt. 13,024009 (2008).
[CrossRef] [PubMed]

K. R. Rau, P. A. Quinto-Su, A. Hellman, and V. Venugopalan, "Pulsed laser microbeam-induced cell lysis: timeresolved imaging and analysis of hydrodynamic effects," Biophys. J. 91,317-329 (2006).
[CrossRef] [PubMed]

Reider, C. L.

V. Magidson, J. Loncarek, P. Hergert, C. L. Reider, and A. Khodjakov,"Laser Microsurgery in the GFP Era: A Cell Biologist???s Perspective," Methods Cell Biol. 82,239-266, (2007).
[PubMed]

Samuel, A. D.

S. H. Chung, D. A. Clark, C. V. Gabel, E. Mazur, and A. D. Samuel,"The role of the AFD neuron in C. elegans thermotaxis analyzed using femtosecond laser ablation," BMC Neuroscience 7, DOI:10.1186/1471-2202-7-30 (2006).
[CrossRef] [PubMed]

Spira, M. E.

H. Erez, G. Malkinson, M. Prager-Khoutorsky, C. I. De Zeeuw, C. C. Hoogenraad, and M. E. Spira, "Formation of microtubule-based traps controls the sorting and concentration of vesicles to restricted sites of regenerating neurons after axotomy," J. Cell Biol. 176,497-507 (2007).
[CrossRef] [PubMed]

Stelzer, E. H. K.

W. Grill, P. Gonczy, E. H. K. Stelzer, and A. A. Hyman "Polarity controls forces governing asymmetric spindle positioning in the Caenorhabditis elegans embryo," Nature 409,630-633 (2001).
[CrossRef] [PubMed]

Tirlapur, U. K.

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

Tokutake, Y.

M. S. Hutson, Y. Tokutake, M.-S. Chang, J.W. Bloor, S. Venakides, D. P. Kiehart, and G. S. Edwards "Forces for morphogenesis investigated with laser-microsurgery and quantitative modeling," Science 300,145-149 (2003).
[CrossRef] [PubMed]

Underwood, J. M.

Venakides, S.

M. S. Hutson, Y. Tokutake, M.-S. Chang, J.W. Bloor, S. Venakides, D. P. Kiehart, and G. S. Edwards "Forces for morphogenesis investigated with laser-microsurgery and quantitative modeling," Science 300,145-149 (2003).
[CrossRef] [PubMed]

Venugopalan, V.

K. R. Rau, P. A. Quinto-Su, A. Hellman, and V. Venugopalan, "Pulsed laser microbeam-induced cell lysis: timeresolved imaging and analysis of hydrodynamic effects," Biophys. J. 91,317-329 (2006).
[CrossRef] [PubMed]

V. Venugopalan, A. GuerraIII, K. Nahen, and A. Vogel, "Role of Laser-Induced Plasma Formation in Pulsed Cellular Microsurgery and Micromanipulation," Phys. Rev. Lett. 88,078103 (2002).
[CrossRef] [PubMed]

Vincent, R.

T. R. Mahoney, Q. Liu, T. Itoh, S. Luo, G. Hadwiger, R. Vincent, Z-W. Wang, M. Fukuda, and M. L. Nonet, "Regulation of synaptic transmission by RAB-3 and RAB-27 in Caenorhabditis elegans," Mol. Biol. Cell 17,2617-2625 (2006).
[CrossRef] [PubMed]

Vogel, A.

A. Vogel, N. Linz, S. Freidank, and G. Paltauf, "Femtosecond-Laser-Induced Nanocavitation in Water: Implications for Optical Breakdown Threshold and Cell Surgery," Phys. Rev. Lett. 100,038102 (2008).
[CrossRef] [PubMed]

V. Venugopalan, A. GuerraIII, K. Nahen, and A. Vogel, "Role of Laser-Induced Plasma Formation in Pulsed Cellular Microsurgery and Micromanipulation," Phys. Rev. Lett. 88,078103 (2002).
[CrossRef] [PubMed]

A. Vogel, M. R. C. Capon, M. N. Asiyo-Vogel, and R. Birngruber, "Intraocular photodisruption with picosecond and nanosecond laser pulses: Tissue effects in cornea, lens, and retina," Invest. Ophthalmol. Visual Sci. 35,3032-3044 (1994).

Wang, Z-W.

T. R. Mahoney, Q. Liu, T. Itoh, S. Luo, G. Hadwiger, R. Vincent, Z-W. Wang, M. Fukuda, and M. L. Nonet, "Regulation of synaptic transmission by RAB-3 and RAB-27 in Caenorhabditis elegans," Mol. Biol. Cell 17,2617-2625 (2006).
[CrossRef] [PubMed]

Watanabe, W.

Weiss, P.

P. Weiss and H. B. Hiscoe,"Experiments on the mechanism of nerve growth," J. Exp. Zool. 107315-395 (1948).
[CrossRef] [PubMed]

Wu, Z.

Z. Wu, A. Ghosh-Roy, M. F. Yanik, J. Z. Zhang, Y. Jin, and A. D. Chisholm, "Caenorhabditis elegans neuronal regeneration is influenced by life stage, ephrin signaling and synaptic branching," PNAS 104,15132-15137 (2007).
[CrossRef] [PubMed]

Yanik, M. F.

Z. Wu, A. Ghosh-Roy, M. F. Yanik, J. Z. Zhang, Y. Jin, and A. D. Chisholm, "Caenorhabditis elegans neuronal regeneration is influenced by life stage, ephrin signaling and synaptic branching," PNAS 104,15132-15137 (2007).
[CrossRef] [PubMed]

M. F. Yanik, H. Cinar, H. N. Cinar, A. Gibby, A. D. Chisholm, Y. Jin, and A. Ben-Yakar, "Nerve regeneration in Caenorhabditis elegans after femtosecond laser axotomy," IEEE J. Quantum Electron. 12,1283-1291 (2006).
[CrossRef]

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]

Zaharieva Maxwell, I.

Zhang, J. Z.

Z. Wu, A. Ghosh-Roy, M. F. Yanik, J. Z. Zhang, Y. Jin, and A. D. Chisholm, "Caenorhabditis elegans neuronal regeneration is influenced by life stage, ephrin signaling and synaptic branching," PNAS 104,15132-15137 (2007).
[CrossRef] [PubMed]

Biophys. J.

K. R. Rau, P. A. Quinto-Su, A. Hellman, and V. Venugopalan, "Pulsed laser microbeam-induced cell lysis: timeresolved imaging and analysis of hydrodynamic effects," Biophys. J. 91,317-329 (2006).
[CrossRef] [PubMed]

BMC Neuroscience

S. H. Chung, D. A. Clark, C. V. Gabel, E. Mazur, and A. D. Samuel,"The role of the AFD neuron in C. elegans thermotaxis analyzed using femtosecond laser ablation," BMC Neuroscience 7, DOI:10.1186/1471-2202-7-30 (2006).
[CrossRef] [PubMed]

Development

J. J. Fernandes and H. Keshishian "Nerve-muscle interactions during flight muscle development in Drosophila," Development,  125,1769-1779 (1998).
[PubMed]

Gene Expr. Patterns

A. Mahr and H. Aberle, "The expression pattern of the Drosophila vesicular glutamate transporter: A marker protein for motoneurons and glutamatergic centers in the brain, Gene Expr. Patterns 6,299-309 (2006).
[CrossRef]

IEEE J. Quantum Electron.

M. F. Yanik, H. Cinar, H. N. Cinar, A. Gibby, A. D. Chisholm, Y. Jin, and A. Ben-Yakar, "Nerve regeneration in Caenorhabditis elegans after femtosecond laser axotomy," IEEE J. Quantum Electron. 12,1283-1291 (2006).
[CrossRef]

Invest. Ophthalmol. Visual Sci.

A. Vogel, M. R. C. Capon, M. N. Asiyo-Vogel, and R. Birngruber, "Intraocular photodisruption with picosecond and nanosecond laser pulses: Tissue effects in cornea, lens, and retina," Invest. Ophthalmol. Visual Sci. 35,3032-3044 (1994).

J. Biomed. Opt.

A. V. Cherian and K. R. Rau, "Pulsed laser-induced damage in rat corneas: time-resolved imaging of physical effects and acute biological response," J. Biomed. Opt. 13,024009 (2008).
[CrossRef] [PubMed]

J. Cell Biol.

H. Erez, G. Malkinson, M. Prager-Khoutorsky, C. I. De Zeeuw, C. C. Hoogenraad, and M. E. Spira, "Formation of microtubule-based traps controls the sorting and concentration of vesicles to restricted sites of regenerating neurons after axotomy," J. Cell Biol. 176,497-507 (2007).
[CrossRef] [PubMed]

J. Exp. Zool.

P. Weiss and H. B. Hiscoe,"Experiments on the mechanism of nerve growth," J. Exp. Zool. 107315-395 (1948).
[CrossRef] [PubMed]

J. Neurosci. Methods

M. L. Nonet, "Visualization of synaptic specializations in live C. elegans with synaptic vesicle protein-GFP fusions," J. Neurosci. Methods 89,33-40 (1999).
[CrossRef] [PubMed]

Methods Cell Biol.

V. Magidson, J. Loncarek, P. Hergert, C. L. Reider, and A. Khodjakov,"Laser Microsurgery in the GFP Era: A Cell Biologist???s Perspective," Methods Cell Biol. 82,239-266, (2007).
[PubMed]

Mol. Biol. Cell

R. V. Barkus, O. Klyachko, D. Horiuchi B. J. Dickson and W. M. Saxton, "Identification of an Axonal Kinesin-3 Motor for Fast Anterograde Vesicle Transport that Facilitates Retrograde Transport of Neuropeptides," Mol. Biol. Cell 19,274-283 (2008).
[CrossRef]

T. R. Mahoney, Q. Liu, T. Itoh, S. Luo, G. Hadwiger, R. Vincent, Z-W. Wang, M. Fukuda, and M. L. Nonet, "Regulation of synaptic transmission by RAB-3 and RAB-27 in Caenorhabditis elegans," Mol. Biol. Cell 17,2617-2625 (2006).
[CrossRef] [PubMed]

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).
[CrossRef]

W. Grill, P. Gonczy, E. H. K. Stelzer, and A. A. Hyman "Polarity controls forces governing asymmetric spindle positioning in the Caenorhabditis elegans embryo," Nature 409,630-633 (2001).
[CrossRef] [PubMed]

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

Neuron

T. Lee and L. Luo, "Mosaic analysis with a repressible cell marker for studies of gene function in neuronal morphogenesis," Neuron 22,451-61 (1999).
[CrossRef] [PubMed]

Opt. Express

Phys. Rev. Lett.

A. Vogel, N. Linz, S. Freidank, and G. Paltauf, "Femtosecond-Laser-Induced Nanocavitation in Water: Implications for Optical Breakdown Threshold and Cell Surgery," Phys. Rev. Lett. 100,038102 (2008).
[CrossRef] [PubMed]

V. Venugopalan, A. GuerraIII, K. Nahen, and A. Vogel, "Role of Laser-Induced Plasma Formation in Pulsed Cellular Microsurgery and Micromanipulation," Phys. Rev. Lett. 88,078103 (2002).
[CrossRef] [PubMed]

M. S. Hutson and X. Ma, "Plasma and cavitation dynamics during pulsed laser microsurgery in vivo," Phys. Rev. Lett. 99,158104 (2007).
[CrossRef] [PubMed]

PNAS

Z. Wu, A. Ghosh-Roy, M. F. Yanik, J. Z. Zhang, Y. Jin, and A. D. Chisholm, "Caenorhabditis elegans neuronal regeneration is influenced by life stage, ephrin signaling and synaptic branching," PNAS 104,15132-15137 (2007).
[CrossRef] [PubMed]

Science

M. S. Hutson, Y. Tokutake, M.-S. Chang, J.W. Bloor, S. Venakides, D. P. Kiehart, and G. S. Edwards "Forces for morphogenesis investigated with laser-microsurgery and quantitative modeling," Science 300,145-149 (2003).
[CrossRef] [PubMed]

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Figures (7)

Fig. 1.
Fig. 1.

Axonal microsurgery of PLM neurons in C. elegans larva with nanosecond laser pulses. The axon is shown prior to (a, c, e) and immediately after surgery (b,d,f). Insets in (b,d,f) show magnified regions around the cut site. Clear breaks can be seen in the axon (b,d). In (f) the extent of damage is extremely small and is only noticeable as a dark band. The proximal end of the axon was also displaced due to the cavitation forces generated during the laser microsurgery. Pulse energy was 0.8 µJ with the laser being operated in single shot mode.

Fig. 2.
Fig. 2.

Manifestions of axonal recovery at different time-points in C. elegans. In (a) vesicle accumulations are seen at 1 hour, in (b) axonal reconnection is seen at 3 hours and in (c) axonal regrowth at 9 hours. Inset in (a) shows a magnified view of the accumulation zone and inset in (b) shows the magnified reconnection zone. Arrow in (c) indicates the growing proximal end, while arrowhead indicates the distal stump.

Fig. 3.
Fig. 3.

Time-resolved imaging of cavitation dynamics in C. elegans. The bubble reaches its maximum size within 20 ns of laser irradiation and the first collapse occurs within ≈30 ns. At later times we observe a rebound and collapse. Arrows indicate the cavitation bubble in different frames.

Fig. 4.
Fig. 4.

Neuronal axotomy in Drosophila 1 st larvae. (a) Two motorneuron bundles prior to axotomy and (b,c) after successive cutting of each bundle. Arrows in (b) and (c) point to the first and second axotomies respectively. Ventral ganglion is at lower left in each image (marked with *). Pulse energy was 1.2 µJ and the laser was operated in single shot mode.

Fig. 5.
Fig. 5.

Time-resolved imaging of cavitation dynamics in Drosophila. The Rmax is seen at 112 ns with collapse at 215 ns.

Fig. 6.
Fig. 6.

Cavitation bubble dynamics in (a) C. elegans and (b) Drosophila. Bubble dynamics are complete within 100 ns in C. elegans and 800 ns in Drosophila.

Fig. 7.
Fig. 7.

(a) Mean areas of GFP::RAB-3 puncta in axons of uncut and axotomized jsIs821. Axons were cut between 10–20 µm from the cell body. The area in (µm2) of GFP::RAB-3 accumulations at the distal and proximal ends of axotomized PLM axons 1 hour after the cut (1.08) is 6 times larger than in uncut axons (0.18) (p<10-4). In cut axons, the mean area at regions away from the cut (0.24) is not statistically different from the uncut axon. Inset shows an image of a cut axon used for quantification, with arrows pointing to accumulations at the proximal and distal ends. * points to cell body. (b) Size distribution of fluorescent accumulations along the axon length. Within 1 µm of the retracted distal and proximal ends of the axon, ≥40% GFP::RAB-3 accumulations are larger than 1 µm2 and more than 20% of the accumulations are between 0.5–1.0 µm2. In uncut axons (Fig. 7(b), inset), fluorescent accumulations with area ≥1 µm2 are not observed while <20% of the puncta are between 0.5–1.0 µm2 in size and the rest are smaller than 0.5 µm2.

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