[Show abstract][Hide abstract] ABSTRACT: Recently developed optogenetics provides a fast, non-invasive, and efficient method for cell activation. However, it is difficult for the optical stimulators used for optogenetics to realize selective multi-site fast activation. In this paper, we developed a random-access photostimulator based on a pair of perpendicularly oriented acousto-optic deflectors. Precise laser targeting in the x-y plane was verified, and the lateral spatial resolution of laser intensity after the objective was measured as ∼1.38 μm. Photostimulaton of ChETA-expressing astrocytes induced reliable inward currents only if the laser beam was directed onto the targeted cell. In the ChR2-expresing neuron, multiple locations along two dendrites were stimulated, and spatiotemporal integration was observed in the soma with fast multi-site activation. These results demonstrated that this random-access photostimulator would be a powerful tool for selective multi-site fast activation. The compact and modular design of this photostimulator makes it easily integrated with different commercial microscopes, and thus widely popularized in many laboratories.
Full-text · Article · Feb 2012 · The Review of scientific instruments
[Show abstract][Hide abstract] ABSTRACT: Light activation and inactivation of neurons by optogenetic techniques has emerged as an important tool for studying neural circuit function. To achieve a high resolution, new methods are being developed to selectively manipulate the activity of individual neurons. Here, we report that the combination of an acousto-optic device (AOD) and single-photon laser was used to achieve rapid and precise spatiotemporal control of light stimulation at multiple points in a neural circuit with millisecond time resolution. The performance of this system in activating ChIEF expressed on HEK 293 cells as well as cultured neurons was first evaluated, and the laser stimulation patterns were optimized. Next, the spatiotemporally selective manipulation of multiple neurons was achieved in a precise manner. Finally, we demonstrated the versatility of this high-resolution method in dissecting neural circuits both in the mouse cortical slice and the Drosophila brain in vivo. Taken together, our results show that the combination of AOD-assisted laser stimulation and optogenetic tools provides a flexible solution for manipulating neuronal activity at high efficiency and with high temporal precision.