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Photoactivation of neurons by laser-generated local heating

机译:激光产生的局部加热对神经元的光激活

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摘要

We present a method for achieving temporally and spatially precise photoactivation of neurons without the need for genetic expression of photosensitive proteins. Our method depends upon conduction of thermal energy via absorption by chemically inert carbon particles and does not require the presence of voltage-gated channels to create transmembrane currents. We demonstrate photothermal initiation of action potentials in Hirudo verbana neurons within an intact ganglion and of transmembrane currents in Xenopus oocytes. Thermal energy is delivered by focused 50 ms, 650 nm laser pulses with total pulse energies between 250 and 3500 μJ. We document an optical delivery system for targeting specific neurons that can be expanded for multiple target sites. Our method achieves photoactivation reliably (70 - 90% of attempts) and can issue multiple pulses (6-9) with minimal changes to cellular properties as measured by intracellular recording. Direct photoactivation presents a significant step towards all-optical analysis of neural circuits in animals such as Hirudo verbana where genetic expression of photosensitive compounds is not feasible.
机译:我们提出了一种方法,可以实现神经元的时间和空间精确的光活化,而无需遗传表达光敏蛋白。我们的方法依靠通过化学惰性碳颗粒的吸收来传导热能,并且不需要电压门控通道的存在来产生跨膜电流。我们展示了完整的神经节内的普通马鞭草神经元的动作电位的光热启动和非洲爪蟾卵母细胞中跨膜电流。通过聚焦的50 ms,650 nm激光脉冲传递热能,总脉冲能量在250至3500μJ之间。我们记录了针对特定神经元的光学传输系统,该系统可以扩展为多个目标站点。我们的方法可靠地实现了光激活(尝试的70-90%),并且可以发出多个脉冲(6-9),而通过细胞内记录所测得的细胞特性变化却很小。直接光激活为全动物神经环路的全光学分析迈出了重要的一步,其中动物马鞭草(Hirudo verbana)不能进行光敏化合物的基因表达。

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