Combined optical tweezers and laser dissector for controlled ablation of functional connections in neural networks.

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  • 04 April 2012
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Regeneration of functional connectivity within a neural network  after different degrees of lesion is of utmost clinical importance. To test pharmacological approaches aimed at recovering from a total  or partial damage of neuronal connections within a circuit, it is  necessary to develop a precise method for controlled ablation of neuronal processes. We combined an UV laser micro-dissector to ablate neural processes in vitro at single neuron and neural  network level with infrared holographic optical tweezers to carry out force spectroscopy measurements. Simultaneous force  spectroscopy, down to the sub-pico-Newton range, was performed  during laser dissection to quantify the tension release in a partially  ablated neurite. Therefore, we could control and measure the  damage inflicted to an individual neuronal process. To characterize the effect of the inflicted injury on network level, changes in  activity of neural sub-populations were monitored with subcellular resolution and overall network activity with high temporal   resolution by concurrent calcium imaging and microelectrode array recording. Neuronal connections have been sequentially ablated  and the correlated changes in network activity traced and mapped. With this unique combination of electrophysiological and optical  tools, neural activity can be and studied and quantified in response  to controlled injury at the subcellular, cellular and network level.