Constructing a low-budget laser axotomy system to study axon regeneration in C. elegans

Area of Science:

• Neuroscience
• Developmental Biology
• Cell Biology

Background:

• Laser axotomy coupled with time-lapse microscopy is a sensitive method for assessing axon regeneration phenotypes in C. elegans.
• The primary limitations of this assay are the high cost and technical expertise associated with traditional laser ablation systems.

Purpose of the Study:

• To describe the construction and implementation of a low-cost, accessible laser axotomy system for C. elegans research.
• To demonstrate that solid-state pulse lasers under $10K can effectively perform laser ablation in transparent biological preparations.

Main Methods:

• A simplified laser ablation system was constructed using a solid-state pulse laser (<$10K) and standard microscopy components.
• An optical path utilizing the microscope condenser's light simplifies alignment, and a dichroic mirror enables simultaneous imaging and ablation.
• Worms were immobilized on agarose pads with anesthetic and microspheres for precise laser axotomy and subsequent time-lapse imaging.

Main Results:

• The constructed system allows for robust laser ablation with a spot size under 1 micrometer.
• Alignment and system construction can be completed within a single day.
• The method enables sensitive assessment of axon regeneration phenotypes in C. elegans.

Conclusions:

• A cost-effective and technically feasible laser axotomy system can be built for C. elegans axon regeneration studies.
• This approach democratizes the use of laser axotomy, broadening its application in neuroscience research.
• The described methodology facilitates the study of nerve repair mechanisms in a transparent model organism.