Gladstone Institutes, United States.
Gladstone Institutes, United States; Department of Neurology, UCSF, United states; Kavli Institute for Fundamental Neuroscience, United States; UCSF Weill Institute for Neurosciences, United States; Department of Physiology, UCSF, United States.
J Neurosci Methods. 2019 Jan 1;311:170-177. doi: 10.1016/j.jneumeth.2018.10.022. Epub 2018 Oct 17.
Intracranial photometry through chronically implanted optical fibers is a widely adopted technique for measuring signals from fluorescent probes in deep-brain structures. The recent proliferation of bright, photo-stable, and specific genetically encoded fluorescent reporters for calcium and for other neuromodulators has greatly increased the utility and popularity of this technique.
Here we describe an open-source, cost-effective, microcontroller-based solution for controlling optical components in an intracranial photometry system and processing the resulting signal.
We show proof-of-principle that this system supports high quality intracranial photometry recordings from dorsal striatum in freely moving mice. A single system supports simultaneous fluorescence measurements in two independent color channels, but multiple systems can be integrated together if additional fluorescence channels are required. This system is designed to work in combination with either commercially available or custom-built optical components. Parts can be purchased for less than one tenth the cost of commercially available alternatives and complete assembly takes less than one day for an inexperienced user.
COMPARISON WITH EXISTING METHOD(S): Currently available hardware draws on a variety of commercial, custom-built, or hybrid elements for both optical and electronic components. Many of these hardware systems are either specialized and inflexible, or over-engineered and expensive.
This open-source system increases experimental flexibility while reducing cost relative to current commercially available components. All software and firmware are open-source and customizable, affording a degree of experimental flexibility that is not available in current commercial systems.
通过慢性植入光纤进行颅内光度测定是一种广泛采用的技术,用于测量深部脑结构中荧光探针的信号。最近,明亮、光稳定且特异性的遗传编码钙和其他神经调质荧光报告基因的大量出现,极大地提高了该技术的实用性和普及性。
本文介绍了一种基于开源、低成本微控制器的解决方案,用于控制颅内光度测定系统中的光学元件并处理产生的信号。
我们证明了该系统支持在自由活动的小鼠背侧纹状体中进行高质量的颅内光度记录。单个系统支持两个独立颜色通道的同时荧光测量,但如果需要更多的荧光通道,则可以将多个系统集成在一起。该系统旨在与商业上可用的或定制的光学组件一起使用。零件的购买成本不到商业上可用替代品的十分之一,对于没有经验的用户,完整的组装过程不到一天。
目前可用的硬件在光学和电子组件方面都采用了各种商业、定制或混合元件。这些硬件系统中的许多都是专业化的和不灵活的,或者是过度设计的和昂贵的。
与当前商业上可用的组件相比,这种开源系统增加了实验的灵活性,同时降低了成本。所有软件和固件都是开源和可定制的,提供了当前商业系统中无法获得的一定程度的实验灵活性。