Brown William G A, Needham Karina, Nayagam Bryony A, Stoddart Paul R
Biotactical Engineering, Faculty of Engineering and Industrial Science, Swinburne University of Technology.
J Vis Exp. 2013 Jul 31(77):50444. doi: 10.3791/50444.
It has been demonstrated in recent years that pulsed, infrared laser light can be used to elicit electrical responses in neural tissue, independent of any further modification of the target tissue. Infrared neural stimulation has been reported in a variety of peripheral and sensory neural tissue in vivo, with particular interest shown in stimulation of neurons in the auditory nerve. However, while INS has been shown to work in these settings, the mechanism (or mechanisms) by which infrared light causes neural excitation is currently not well understood. The protocol presented here describes a whole cell patch clamp method designed to facilitate the investigation of infrared neural stimulation in cultured primary auditory neurons. By thoroughly characterizing the response of these cells to infrared laser illumination in vitro under controlled conditions, it may be possible to gain an improved understanding of the fundamental physical and biochemical processes underlying infrared neural stimulation.
近年来已证实，脉冲红外激光可用于激发神经组织中的电反应，而无需对目标组织进行任何进一步的修饰。体内多种外周和感觉神经组织中均有红外神经刺激的报道，其中对听神经神经元的刺激尤为受关注。然而，尽管已证明红外神经刺激在这些情况下有效，但目前对红外光引起神经兴奋的机制尚不清楚。本文介绍的方案描述了一种全细胞膜片钳方法，旨在促进对培养的原代听觉神经元中红外神经刺激的研究。通过在受控条件下全面表征这些细胞在体外对红外激光照射的反应，有可能更好地理解红外神经刺激背后的基本物理和生化过程。
