Sacconi Leonardo, Mapelli Jonathan, Gandolfi Daniela, Lotti Jacopo, O'Connor Rodney P, D'Angelo Egidio, Pavone Francesco S
European Laboratory for Nonlinear Spectroscopy, University of Florence, Italy.
Opt Express. 2008 Sep 15;16(19):14910-21.
One of the main challenges in understanding the central nervous system is to measure the network dynamics of neuronal assemblies, while preserving the computational role of individual neurons. However, this is not possible with current techniques. In this work, we combined the advantages of second-harmonic generation (SHG) with a random access (RA) excitation scheme to realize a new microscope (RASH) capable of optically recording fast membrane potential events occurring in a wide-field of view. The RASH microscope, in combination with bulk loading of tissue with FM4-64 dye, was used to simultaneously record electrical activity from clusters of Purkinje cells in acute cerebellar slices. Complex spikes, both synchronous and asynchronous, were optically recorded simultaneously across a given population of neurons. Spontaneous electrical activity was also monitored simultaneously in pairs of neurons, where action potentials were recorded without averaging across trials. These results show the strength of this technique in describing the temporal dynamics of neuronal assemblies, opening promising perspectives in understanding the computations of neuronal networks.
理解中枢神经系统的主要挑战之一是测量神经元集合的网络动态，同时保留单个神经元的计算作用。然而，目前的技术无法做到这一点。在这项工作中，我们将二次谐波产生（SHG）的优势与随机访问（RA）激发方案相结合，实现了一种新型显微镜（RASH），能够光学记录在宽视野中发生的快速膜电位事件。RASH显微镜与用FM4-64染料对组织进行批量加载相结合，用于同时记录急性小脑切片中浦肯野细胞簇的电活动。在给定的神经元群体中，同步和异步的复合尖峰都被光学同时记录下来。还同时监测了成对神经元的自发电活动，其中动作电位是在不进行试验平均的情况下记录的。这些结果显示了该技术在描述神经元集合的时间动态方面的优势，为理解神经网络的计算开辟了广阔前景。
