Novel Nano-pipette for imaging of deep cortical layers and deep brain structures

描述(申请人提供)：在这里，我们建议开发一种多功能纳米吸管(NP)，用于活体内窥镜成像，并结合来自皮质深层或脑深部结构的电生理记录。利用这一NP，我们计划研究跨皮质和来自大脑深部核团的信息处理的神经元机制。目前，大鼠桶状皮质和行为猴的视皮层的功能图谱已经很好地建立起来，并可以使用功能成像技术在2D上显示，例如具有高空间分辨率的本征信号光学成像(OI-IS)，或具有高时空分辨率的电压敏感染料成像(VSDI)。然而，由于现代活体成像技术无法从大脑皮层深层分辨出高时空分辨率的神经元活动，3D成像功能图尚未完全实现。出于同样的原因，目前还不可能使用VSDI或OI-IS对大脑深层结构进行成像。在这里，我们建议通过结合纳米技术和活体实时成像技术领域的最新发展来解决这个问题。最近，Zeev Zalevsky教授和他的实验室开发了一种技术，能够产生类似于光子晶体光纤(PCF)的光纤，具有内部金属线。这些纤维是由具有所需横截面的锥形二氧化硅预制件产生的，金属丝被预先插入其中。这种在不失去内部几何结构的情况下获得的预制件的锥形，产生了将光和电信号传输到合适的传感器的短而薄的纳米吸管。此外，扎列夫斯基实验室已经开发出算法来放大这种系统固有的低分辨率。利用这项技术和专业知识，结合Hamutal斯洛伐in博士的成像专业知识，我们建议制造包含光纤和电纤维的锥形预制件(特定目标1)，并测试其在体外和体内小型人造目标上的适用性(特定目标2)。最后，我们将通过功能成像，即OI-IS和VSDI(特定目标3)，在麻醉大鼠的桶状皮质和脑深部核团上进行活体测试，以确定该设备是否可以用于脑成像。如果成功，我们的纳米吸管有望彻底改变现有的成像能力，并将使人们能够对视觉感知和更高认知功能背后的神经机制有新的见解。与公共健康相关：我们建议开发一种纳米管(探头)，它将被插入大脑的皮质表面，并将能够从皮质深层或大脑深层核团同时进行成像和电生理记录。如果成功，该项目的结果将彻底改变现有的成像能力，并将使人们能够对潜在的视觉感知和更高认知功能的神经机制有新的见解。

DESCRIPTION (provided by applicant): Here we propose to develop a multi-functional nano-pipette (NP) for in vivo endoscopic imaging, combined with electrophysiological recording from deep cortical layers or deep brain structures. Using this NP, we plan to study neuronal mechanisms of information processing across cortical layers and from deep brain nuclei. Functional maps in the barrel cortex of rats and visual cortex of behaving monkeys are well established nowadays and can be demonstrated in 2D, using functional imaging techniques such as optical imaging of intrinsic signals (OI-IS), with high spatial resolution, or voltage-sensitive dye imaging (VSDI) with high spatio-temporal resolution. However, imaging functional maps in 3D has not yet been fully achieved due to the inability of modern in vivo imaging techniques to resolve neuronal activity at high spatio-temporal resolution from deep layers of the cerebral cortex. For the same reasons, imaging from deep brain structures using VSDI or OI-IS is not possible currently. Here we propose to address this problem by combining recent developments in the field on nanotechnology and in real time imaging techniques in vivo. Recently, Prof. Zeev Zalevsky and his laboratory have developed the technology that enables generation of fibers similar to photonic crystal fibers (PCF), having internal metal wires. Those fibers are generated by tapering silica pre-forms with desired cross section into which the wires are pre-inserted. This tapering of the pre-form, obtained without losing its internal geometry, produces short, thin nano-pipettes that transmit light and electrical signals to appropriate sensors. Furthermore, the Zalevsky laboratory has developed algorithms to amplify the low resolution inherent in such a system. Using this technology and expertise, in combination with Dr. Hamutal Slovin's imaging expertise, we propose to fabricate a tapered pre-form containing optical and electrical fibers (Specific Aim #1) and test its applicability on small artificial targets in vitro and in vivo (Specific Aim #2). Finally, we will determine whether this device can be used for brain imaging by testing it in vivo on the barrel cortex and deep brain nuclei of anesthetized rats through functional imaging, namely OI-IS and VSDI (Specific Aim #3). If successful, our nano-pipette is expected to revolutionize existing imaging capabilities and will enable new insights into the neural mechanisms underlying visual perception and higher cognitive functions. PUBLIC HEALTH RELEVANCE: We propose to develop a nanopipette (probe), which will be inserted into the cortical surface of the brain and will enable simultaneous imaging and electrophysiological recording from deep cortical layers or deep brain nuclei. If successful, the outcomes of this project will revolutionize existing imaging capabilities and will enable new insights into the neural mechanisms underlying visual perception and higher cognitive functions.