Institute for Physical Science and Technology, University of Maryland, College Park, Maryland 20742.
Fraunhofer USA Center Mid-Atlantic, Riverdale, Maryland 20737.
eNeuro. 2024 Oct 16;11(10). doi: 10.1523/ENEURO.0079-24.2024. Print 2024 Oct.
Two-photon calcium imaging allows for the activity readout of large populations of neurons at single cell resolution in living organisms, yielding new insights into how the brain processes information. Holographic optogenetics allows us to trigger activity of this population directly, raising the possibility of injecting information into a living brain. Optogenetic triggering of activity that mimics "natural" information, however, requires identification of stimulation targets based on real-time analysis of the functional network. We have developed NeuroART (Neuronal Analysis in Real Time), software that provides real-time readout of neuronal activity integrated with downstream analysis of correlations and synchrony and of sensory metadata. On the example of auditory stimuli, we demonstrate real-time inference of the contribution of each neuron in the field of view to sensory information processing. To avoid the limitations of microscope hardware and enable collaboration of multiple research groups, NeuroART taps into microscope data streams without the need for modification of microscope control software and is compatible with a wide range of microscope platforms. NeuroART also integrates the capability to drive a spatial light modulator (SLM) for holographic photostimulation of optimal stimulation targets, enabling real-time modification of functional networks. Neurons used for photostimulation experiments were extracted from Sprague Dawley rat embryos of both sexes.
双光子钙成像允许在活生物体中单细胞分辨率对大量神经元的活动进行读出，为大脑如何处理信息提供了新的见解。全息光遗传学使我们能够直接触发该群体的活动，从而有可能将信息注入活体大脑。然而，要模拟“自然”信息的光遗传学触发活动，就需要基于功能网络的实时分析来确定刺激目标。我们开发了 NeuroART（实时神经元分析）软件，该软件可实时读取神经元活动，并对相关性和同步性以及感官元数据进行下游分析。我们以听觉刺激为例，实时推断视野中每个神经元对感官信息处理的贡献。为了避免显微镜硬件的限制并使多个研究小组能够协作，NeuroART 无需修改显微镜控制软件即可利用显微镜数据流，并与多种显微镜平台兼容。NeuroART 还集成了驱动空间光调制器 (SLM) 的功能，用于全息光刺激最佳刺激目标，从而能够实时修改功能网络。用于光刺激实验的神经元取自雄性和雌性 Sprague Dawley 大鼠胚胎。
