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FIGURE 1. From: A Tribute to Dr. David Kline: A New Approach to an Old Peripheral Nerve Problem--Splicing Instead of Regenerating Disrupted Axons.

In the direct repair scheme recently proposed, novel microsurgical instruments directly manipulate individual axons in an injured nerve. A microcutting tool cuts each axon to remove damaged segments, leaving a discrete, resealing stump. Electrostatic forces are then used to align and juxtapose individual axonal segments in preparation for rejoining. Electrofusion is then applied to the juxtaposed axons, splicing the segments together and leading to reconstitution of a functional axon. A, schematic illustrating this technique. B, a microcutting tool is positioned over a single axon. BC, the axon has been cut (arrow). The tool is able to deliver neat and precise cuts to individual axons, leaving a discrete end that can reseal. Following the cutting, intact axonal segments are aligned and juxtaposed with each other ahead of the splicing step. D and E, the stretching and aligning of axons between electrodes due to the application of dielectrophoretic force (arrows). Using electrofusion, the juxtaposed axons are joined together to form a single intact unit. F and G, the fusion of 2 axons allows the fluorescent protein in 1 axon to diffuse into its previously fluorescent-free partner (double arrows). Retention of the fluorescence indicates that membrane continuity is maintained.

Wesley Chang, et al. Neurosurgery. ;65(4 Suppl):A52-A54.

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