Abstract:
Behaviour is the ultimate output of neural circuit computations, and therefore its analysis is a cornerstone of neuroscience research. However, every animal and experimental paradigm requires different illumination conditions to capture and, in some cases, manipulate specific behavioural features. This means that researchers often develop, from scratch, their own solutions and experimental set-ups. Here, we present OptoPi, an open source, affordable (∼ £600), behavioural arena with accompanying multi-animal tracking software. The system features highly customisable and reproducible visible and infrared illumination and allows for optogenetic stimulation. OptoPi acquires images using a Raspberry Pi camera, features motorised LED-based illumination, Arduino control, as well as irradiance monitoring to fine-tune illumination conditions with real time feedback. Our open-source software (BioImageProcessing) can be used to simultaneously track multiple unmarked animals both in on-line and off-line modes. We demonstrate the functionality of OptoPi by recording and tracking under different illumination conditions the spontaneous behaviour of larval zebrafish as well as adult Drosophila flies and their first instar larvae, an experimental animal that due to its small size and transparency has classically been hard to track. Further, we showcase OptoPi\'s optogenetic capabilities through a series of experiments using transgenic Drosophila larvae.
摘要:
行为是神经回路计算的最终输出,因此,它的分析是神经科学研究的基石。然而,每个动物和实验范式都需要不同的光照条件来捕捉,在某些情况下,操纵特定的行为特征。这意味着研究人员经常开发,从头开始,他们自己的解决方案和实验设置。这里,我们展示了OptoPi,一个开源,负担得起的(600英镑),带有多动物跟踪软件的行为竞技场。该系统具有高度可定制和可重复的可见和红外照明,并允许光遗传刺激。OptoPi使用RaspberryPi相机获取图像,具有电动LED照明,Arduino控制,以及辐照度监测,以实时反馈微调照明条件。我们的开源软件(BioImageProcessing)可用于在在线和离线模式下同时跟踪多个未标记的动物。我们通过在不同光照条件下记录和跟踪幼体斑马鱼以及成年果蝇及其第一龄幼虫的自发行为来证明OptoPi的功能,一种实验动物,由于它的小尺寸和透明度一直很难追踪。Further,我们通过一系列使用转基因果蝇幼虫的实验展示了OptoPi的光遗传学能力。