Harrison Thomas C, Sigler Albrecht, Murphy Timothy H
Kinsmen Laboratory, Department of Psychiatry, 2255 Wesbrook Mall, University of British Columbia, Vancouver, BC, Canada V6T 1Z3.
J Neurosci Methods. 2009 Sep 15;182(2):211-8. doi: 10.1016/j.jneumeth.2009.06.021. Epub 2009 Jun 24.
We describe a simple and low-cost system for intrinsic optical signal (IOS) imaging using stable LED light sources, basic microscopes, and commonly available CCD cameras. IOS imaging measures activity-dependent changes in the light reflectance of brain tissue, and can be performed with a minimum of specialized equipment. Our system uses LED ring lights that can be mounted on standard microscope objectives or video lenses to provide a homogeneous and stable light source, with less than 0.003% fluctuation across images averaged from 40 trials. We describe the equipment and surgical techniques necessary for both acute and chronic mouse preparations, and provide software that can create maps of sensory representations from images captured by inexpensive 8-bit cameras or by 12-bit cameras. The IOS imaging system can be adapted to commercial upright microscopes or custom macroscopes, eliminating the need for dedicated equipment or complex optical paths. This method can be combined with parallel high resolution imaging techniques such as two-photon microscopy.
我们描述了一种用于固有光学信号(IOS)成像的简单且低成本的系统,该系统使用稳定的LED光源、基本显微镜和常用的CCD相机。IOS成像可测量脑组织光反射率中与活动相关的变化,并且只需最少的专用设备即可进行。我们的系统使用可安装在标准显微镜物镜或视频镜头上的LED环形灯,以提供均匀且稳定的光源,在40次试验平均得到的图像中,其波动小于0.003%。我们描述了急性和慢性小鼠制备所需的设备和手术技术,并提供了可根据廉价8位相机或12位相机拍摄的图像创建感觉表征图谱的软件。该IOS成像系统可适配于商用直立显微镜或定制宏观显微镜,无需专用设备或复杂的光路。此方法可与双光子显微镜等并行高分辨率成像技术相结合。