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Bioengineering

De bouw van een Low-budget Laser axotomy Systeem om Axon Regeneration Study in C. elegans doi: 10.3791/3331 Published: November 15, 2011

Materials

Name Company Catalog Number Comments
Name of the reagent Company Catalogue number Comments (optional)
535 nm Laser Crylas FDSS532Q3 (TEEM, Crystal, and CRC also offer comparable lasers)
All optics and hardware (Thor offers comparable optics and hardware)
SUPREMA Optical Mount, 1.0-in diameter Newport Corp. SS100-F2KN 2
1" diameter post, 1" height Newport Corp. PS-1 1
1" diameter post fork Newport Corp. PS-F 1
Achromatic Zero-Order Wavel Plate, ½ Wave Retardation, 400-700nm Newport Corp. 10RP52-1 1
Rotation Stage, 1" Aperature Newport Corp. RSP-1T 1
1" diameter post, 1" height Newport Corp. PS-1 1
1" diameter post fork Newport Corp. PS-F 1
Glan-Laser Calcite Polarizer, 430-700nm Newport Corp. 10GL08AR.14 1
Polarizer Rotation Mount Newport Corp. RM25A 1
¼" spacer for 1" diameter post Newport Corp. PS-0.25 1
½" height, 1"diameter post Newport Corp. PS-0.5 1
Fork, 1" diameter post Newport Corp. PS-F 1
Beam Dump Newport Corp. PL15 1
Microscope Objective Lens Mount Newport Corp. LH-OBJ1 1
1" diameter post, 1" height Newport Corp. PS-1 1
1" diameter post fork Newport Corp. PS-F 1
High-Energy Nd:YAG Laser Mirror, 25.4 mm Diameter, 45°, 532 nm Newport Corp. 10Q20HE.2 4
SUPREMA Optical Mount, 1.0 inch diameter, clearance mounting hole Newport Corp. SN100C-F 2
High Precision Knob Adjustment Screw, 12.7mm travel, 100TPI Newport Corp. AJS100-0.5K 4
Thread adaptor, ¼-20 male, 8-32 female Newport Corp. SS-1-B 2
Rod Clamp for 1.5 inch diameter rod Newport Corp. 340-RC 2
Rod, 14 inch (35.5 cm) tall Newport Corp. 40 1
Rail carrier for X26, square 40mm length Newport Corp. CN26-40 4
Steel Rail, 384mm (15") length Newport Corp. X26-384 1
Rod Platform for 1.5 inch diameter rod Newport Corp. 300-P 2
Rod, 14 inch (35.5 cm) tall Newport Corp. 40 2
Plano-Concave Lens, 12.7mm diameter, -25mm EFL, 430-700nm Newport Corp. KPC025AR.14 2
Plano-Convex Lens, 25.4mm diameter, 50.2mm EFL, 430-700nm Newport Corp. KPX082AR.14 1
Plano-Convex Lens, 25.4mm diameter, 62.9mm EFL, 430-700nm Newport Corp. KPX085AR.14 1
Fixed Lens Mount, 0.5" diameter, 1.0" Axis height Newport Corp. LH-0.5 2
Fixed Lens Mount, 1.0" diameter, 1.0" Axis height Newport Corp. LH-1 2
Microspheres 0.1um Polysciences, Inc. 00876
Agarose Research Products International Corp. A20090 EEO matters
Muscimol Sigma-Aldrich M1523

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References

  1. Hammarlund, M., Nix, P., Hauth, L., Jorgensen, E. M., Bastiani, M. Axon regeneration requires a conserved MAP kinase pathway. Science. 323, 802-806 (2009).
  2. Steinmeyer, J. D. Construction of a femtosecond laser microsurgery system. Nature protocols. 5, 395-407 (2010).
  3. Wu, Z. Caenorhabditis elegans neuronal regeneration is influenced by life stage, ephrin signaling, and synaptic branching. Proc Natl Acad. 104, 15132-15137 (2007).
  4. Gilleland, C. L., Rohde, C. B., Zeng, F., Yanik, M. F. Microfluidic immobilization of physiologically active Caenorhabditis elegans. Nature protocols. 5, 1888-1902 (2010).
  5. Raabe, I., Vogel, S. K., Peychl, J., Tolic-Norrelykke, I. M. Intracellular nanosurgery and cell enucleation using a picosecond laser. J. Microsc. 234, 1-8 (2009).
  6. Hutson, M. S., Ma, X. Plasma and cavitation dynamics during pulsed laser microsurgery in vivo. Physical review letters. 99, 158104-158104 (2007).
  7. Venugopalan, V., Guerra, A. 3rd, Nahen, K., Vogel, A. Role of laser-induced plasma formation in pulsed cellular microsurgery and micromanipulation. Physical review letters. 88, 078103-078103 (2002).
  8. Bourgeois, F., Ben-Yakar, A. Femtosecond laser nanoaxotomy properties and their effect on axonal recovery in C. elegans. Opt Express. 16, 5963-5963 (2008).
  9. O'Brien, G. S., Rieger, S., Martin, S. M., Cavanaugh, A. M., Portera-Cailliau, C., Sagasti, A. Two-photon axotomy and time-lapse confocal imaging in live zebrafish embryos. J. Vis. Exp. (24), e1129-e1129 (2009).
  10. Tsai, P. S. Plasma-mediated ablation: an optical tool for submicrometer surgery on neuronal and vascular systems. Curr. Opin. Biotechnol. 20, 90-99 (2009).
  11. Chung, S. H., Clark, D. A., Gabel, C. V., Mazur, E., Samuel, A. D. The role of the AFD neuron in C. elegans thermotaxis analyzed using femtosecond laser ablation. BMC Neurosci. 7, 30-30 (2006).
  12. Shen, N. Ablation of cytoskeletal filaments and mitochondria in live cells using a femtosecond laser nanoscissor. Mech. Chem. Biosyst. 2, 17-25 (2005).
  13. Rao, G. N., Kulkarni, S. S., Koushika, S. P., Rau, K. R. In vivo nanosecond laser axotomy: cavitation dynamics and vesicle transport. Opt. Express. 16, 9884-9894 (2008).
  14. Rohde, C. B., Yanik, M. F. Subcellular in vivo time-lapse imaging and optical manipulation of Caenorhabditis elegans in standard multiwell plates. Nat. Commun. 2, 271-271 (2011).
De bouw van een Low-budget Laser axotomy Systeem om Axon Regeneration Study in<em> C. elegans</em
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Cite this Article

Williams, W., Nix, P., Bastiani, M. Constructing a Low-budget Laser Axotomy System to Study Axon Regeneration in C. elegans. J. Vis. Exp. (57), e3331, doi:10.3791/3331 (2011).More

Williams, W., Nix, P., Bastiani, M. Constructing a Low-budget Laser Axotomy System to Study Axon Regeneration in C. elegans. J. Vis. Exp. (57), e3331, doi:10.3791/3331 (2011).

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