Laser axotomy was performed as previously described in Byrne et al., 2014 [46 (link)]. L4 animals were immobilized using 0.05 μm polystyrene beads (Polybead Microspheres, Polysciences Cat #08691–10) or in 0.2mM Levamisole (Sigma) on a pad of 3% agarose dissolved in M9 buffer on a glass slide. Worms were visualized using a Nikon Eclipse 80i microscope with a 100x Plan Apo VC lens (1.4 NA). Fluorescently-labeled D-type motor neuron commissures were targeted at the dorsoventral midline using a 435 nm Micropoint laser with 10 pulses at 20 Hz. In all cases no more than four of the seven posterior commisures were cut per animal to minimize possible adverse locomotion or behavioral effects. Animals were recovered to NGM plates seeded with OP50 and allowed to recover.
Plan apo vc lens
Manufactured by Nikon
The Plan Apo VC lens from Nikon is a high-performance optical lens designed for laboratory and research applications. It features a plan-apochromatic design that ensures excellent image quality and resolution across the entire field of view. The lens is also equipped with Vibration Compensation (VC) technology, which helps to reduce the effects of hand shake or other external vibrations, providing stable and clear images. The details of the lens's intended use or performance characteristics are not available in this factual and unbiased description.
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2 protocols using plan apo vc lens
1
Laser Axotomy of D-type Motor Neurons
2
Laser Axotomy of Motor Neuron Commissures
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Laser axotomy was performed as previously described in Byrne et al. 2011 (link). L4 animals were immobilized using 0.05 µm polystyrene beads (Polybead Microspheres, Polysciences Cat #08691-10) or in 0.2mM Levamisole (Sigma) on a pad of 3% agarose dissolved in M9 buffer on a glass slide. Worms were visualized using a Nikon Eclipse 80i microscope with a 100x Plan Apo VC lens (1.4 NA). Fluorescently-labeled D-type motor neuron commissures were targeted at the dorsoventral midline using a 435 nm Micropoint laser with 10 pulses at 20 Hz. In all cases no more than four of the seven posterior commisures were cut per animal to minimize possible adverse locomotion or behavioral effects. Animals were recovered to NGM plates seeded with OP50 and allowed to recover.
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