Research Institute for Electronic Science, Hokkaido University, N20, W10, Sapporo, Hokkaido 001-0020, Japan.
Neurosci Res. 2012 Aug;73(4):341-7. doi: 10.1016/j.neures.2012.05.007. Epub 2012 May 28.
Optogenetic tools, such as channelrhodopsin2 (ChR2), have enabled the behavior of whole organisms by light-mediated manipulation of neuronal activities. Fluorescent indicators have been used to aid in the understanding of what is happening in living cells. To date, optogenetic stimulation and imaging acquisition were sequentially performed during detector "live time." However, there is a problem with interrupting acquisition time sequences because such stimulation invades the time territory of fluorescent imaging. Here, our purpose was to show that optogenetic stimulation can be performed within the "dead time" of the charge-coupled device camera, the short interval of data transfer between frames. We show the kinetic measurement of Ca(2+) dynamics in neuron-like cells upon ChR2 stimulation, by which we reveal the biphasic property of the Ca(2+) increase in response to optical stimulation.
光遗传学工具，如通道视紫红质 2（ChR2），通过光介导的神经元活动操纵，使整个生物体的行为成为可能。荧光指示剂被用于帮助理解活细胞中发生的事情。迄今为止，光遗传学刺激和成像采集在探测器“实时”期间顺序进行。然而，由于这种刺激侵犯了荧光成像的时间范围，因此存在中断采集时间序列的问题。在这里，我们的目的是表明光遗传学刺激可以在电荷耦合器件（CCD）相机的“死时间”内进行，这是帧与帧之间数据传输的短暂间隔。我们展示了在 ChR2 刺激下神经元样细胞中 Ca(2+)动力学的动力学测量，通过该测量，我们揭示了光刺激响应中 Ca(2+)增加的双相特性。
