Development of photoconductive stimulation technology for long-term interfacing with living neurons.

传统上，认知脑功能的研究一直是通过使用人类受试者或动物模型来进行的。然而，过去几十年的技术进步现在允许我们使用体外神经元系统以高分辨率调查负责高级脑功能的潜在机制。理解和发展研究神经元活动模式的技术有许多应用：从直接促进我们对复杂动力系统的理解到药物筛选设备的创建和神经病理条件的建模。这项研究的关键是设备和系统的发展，这些设备和系统允许人们同时刺激和记录活的神经网络。其中一项技术，光导刺激，是我和我的同事在加州大学圣地亚哥分校的Yukiko Goda实验室发明的。基于在可见光照射下硅的导电性会发生变化的原理，神经元首先生长在专门制造的硅晶片表面。然后，通过光瞄准它们，它们可以被单独触发，以用户定义的频率或以特定的非随机模式发射。结合钙或电压敏感染料成像的神经元网络的电活动，同时与数百个神经元双向通信已成为现实。本提案旨在通过以下方式扩展光导刺激技术，以实现以下具体目标：******1)开发长期神经元-硅界面室和刺激方法，用于与活神经元培养物扩展甚至永久界面。***2)开发原位光学活性记录设备和协议，以促进神经元活动的扩展高分辨率记录和分析。******中心假设：通过使用基于光学的神经元/硅接口，我们已经建立了一种非侵入性的方法来获得活神经元网络的读/写能力。通过开发维持这种功能的长期受控环境，我们可以研究环境和遗传操纵对脑回路的影响的关键问题，并获得准确分析活神经元网络复杂系统动力学所需的高空间和时间分辨率数据。******我们相信，一个可行的长期神经元接口平台的发展不仅会给我们对大脑功能和复杂动力系统的理解带来直接的收益，而且还将有许多与其他研究和商业目标相结合的应用

Traditionally, the study of cognitive brain function has been approached through the use of either human subjects or animal models. However, technological advances from the last few decades now allow us to use in vitro neuronal systems to investigate in high resolution the underlying mechanisms responsible for higher level brain functions. Understanding and developing technology to investigate neuronal activity patterns has many applications: from directly facilitating our understanding of complex dynamical systems through to the creation of drug screening devices and modeling of neuropathological conditions. Essential to this research has been the development of devices and systems that allow one to both stimulate and record from living neuronal networks. One such technology, photoconductive stimulation, was invented by my colleagues and I in the lab of Yukiko Goda at UCSD. Based on the principle that the conductivity of silicon will change when illuminated by visible light, neurons are first grown on the surface of specifically manufactured silicon wafers. Then, by optically targeting them with light, they can be individually triggered to fire at a user-defined frequency or in a specific non-random pattern. Combined with calcium- or voltage-sensitive dye imaging of the neuronal network's electrical activity, simultaneous two-way communication with hundreds of neurons has become a reality. This proposal seeks to extend the photoconductive stimulation technology in the following ways, for the following specific goals:******1) Development of a long-term neuronal-silicon interface chamber and stimulation methodology for extended or even permanent interfacing with living neuronal cultures.***2) Development of the in situ optical activity recording devices and protocols, to facilitate extended high resolution recording and analysis of neuronal activity.******Central Hypothesis: Through the use of an optically-based neuronal/silicon interface, we have established a non-invasive methodology to gain read/write capability with living neuronal networks. By developing a long-term controlled environment that maintains this functionality, we can pursue critical questions as to the effect of environmental and genetic manipulation of brain circuits, as well as obtain high spatial- and temporal-resolution data necessary to accurately analyze the complex system dynamics of living neuronal networks.******We believe the development of a viable long-term neuronal interface platform will not only bring immediate gains in our understanding of brain function and complex dynamical systems, but will also have numerous applications for integrating with other research and commercial goals.**