✓Set up and calibrate a MicroPoint pulsed laser ablation system for C. elegans
✓Immobilize live nematodes for precise in vivo laser targeting
✓Perform controlled axotomy to sever labeled neuronal processes
✓Assess neuronal regeneration following laser-induced injury
Protocol
Biopharma Insights A protocol to cut neurons in C. elegans with a MicroPoint pulsed laser is presented. We describe setting up the system, immobilizing worms, and severing labeled neurons. Advantages include a relatively low-cost system and the ability to sever neuronal processes or ablate cells in vivo.
Difficulty
advanced
Total time
~1–2 hours per experimental session (setup + 5–10 worms)
Model organism
Caenorhabditis elegans
Steps
1
Set up laser ablation system and optics
Assemble and initialize the MicroPoint pulsed laser system, align optical components, and verify beam path to the microscope objective.
▶ 01:02
2
Focus laser in Z-plane (axial depth)
Adjust objective focus along the vertical axis to position the laser focal point at the target neuronal structure within the living worm.
▶ 02:17
3
Focus laser in X-Y plane (lateral position)
Center the laser spot on the target neuron in the horizontal plane using stage micromanipulation and real-time visualization.
▶ 04:12
4
Immobilize worms for laser ablation
Restrain live C. elegans on a slide or in a microfluidic chamber to prevent movement during laser targeting and axotomy.
▶ 05:00
5
Perform laser axotomies on target neurons
Deliver pulsed laser energy to sever the focused neuronal process, confirming physical transection of the labeled axon.
▶ 06:05
6
Score neurons for regeneration capacity
Assess and document neuronal regrowth at the axotomy site at defined time points post-injury to evaluate regenerative potential.
▶ 07:57
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