Barral Jérémie, Reyes Alex D
Center for Neural Science, New York University, New York, USA.
Bio Protoc. 2017 Jun 20;7(12). doi: 10.21769/BioProtoc.2335.
We studied a network of cortical neurons in culture and developed an innovative optical device to stimulate optogenetically a large neuronal population with both spatial and temporal precision. We first describe how to culture primary neurons expressing channelrhodopsin. We then detail the optogenetic setup based on the workings of a fast Digital Light Processing (DLP) projector. The setup is able to stimulate tens to hundreds neurons with independent trains of light pulses that evoked action potentials with high temporal resolution. During photostimulation, network activity was monitored using patch-clamp recordings of up to 4 neurons. The experiment is ideally suited to study recurrent network dynamics or biological processes such as plasticity or homeostasis in a network of neurons when a sub-population is activated by distinct stimuli whose characteristics (correlation, rate, and, size) were finely controlled.
我们研究了培养的皮质神经元网络，并开发了一种创新的光学装置，以时空精确性对大量神经元群体进行光遗传学刺激。我们首先描述如何培养表达通道视紫红质的原代神经元。然后详细介绍基于快速数字光处理（DLP）投影仪工作原理的光遗传学设置。该设置能够用独立的光脉冲序列刺激数十到数百个神经元，这些光脉冲序列能以高时间分辨率诱发动作电位。在光刺激期间，使用膜片钳记录多达4个神经元来监测网络活动。当一个亚群由特征（相关性、频率和大小）得到精细控制的不同刺激激活时，该实验非常适合研究神经元网络中的递归网络动力学或生物过程，如可塑性或稳态。
