Department of Power Mechanical Engineering, Institute of Biotechnology, Brain Research Center, Institute of Photonics Technologies, Department of Electrical Engineering, and Institute of NanoEngineering and MicroSystems, National Tsing Hua University, Hsinchu 30013, Taiwan.
Proc Natl Acad Sci U S A. 2014 Apr 8;111(14):5367-72. doi: 10.1073/pnas.1400997111. Epub 2014 Mar 24.
We present an automated laser tracking and optogenetic manipulation system (ALTOMS) for studying social memory in fruit flies (Drosophila melanogaster). ALTOMS comprises an intelligent central control module for high-speed fly behavior analysis and feedback laser scanning (∼40 frames per second) for targeting two lasers (a 473-nm blue laser and a 593.5-nm yellow laser) independently on any specified body parts of two freely moving Drosophila adults. By using ALTOMS to monitor and compute the locations, orientations, wing postures, and relative distance between two flies in real time and using high-intensity laser irradiation as an aversive stimulus, this laser tracking system can be used for an operant conditioning assay in which a courting male quickly learns and forms a long-lasting memory to stay away from a freely moving virgin female. With the equipped lasers, channelrhodopsin-2 and/or halorhodopsin expressed in selected neurons can be triggered on the basis of interactive behaviors between two flies. Given its capacity for optogenetic manipulation to transiently and independently activate/inactivate selective neurons, ALTOMS offers opportunities to systematically map brain circuits that orchestrate specific Drosophila behaviors.
我们展示了一个自动化的激光跟踪和光遗传学操纵系统（ALTOMS），用于研究果蝇（Drosophila melanogaster）的社交记忆。ALTOMS 包括一个智能中央控制模块，用于高速飞行行为分析和反馈激光扫描（每秒约 40 帧），以独立瞄准两只自由移动的果蝇的两个指定身体部位的两个激光器（473nm 蓝色激光和 593.5nm 黄色激光）。通过使用 ALTOMS 实时监测和计算两只果蝇的位置、方向、翅膀姿势和相对距离，并使用高强度激光照射作为厌恶刺激，这个激光跟踪系统可用于操作性条件反射测定，其中求爱雄性会迅速学习并形成持久的记忆，远离自由移动的处女雌性。通过配备的激光器，可以根据两只果蝇之间的互动行为，触发特定神经元中的通道视紫红质-2 和/或卤化视蛋白。鉴于其对选择性神经元进行瞬时和独立激活/失活的光遗传学操纵能力，ALTOMS 为系统地绘制协调特定果蝇行为的大脑回路提供了机会。
