High-speed volumetric imaging of neuronal network activity at depth using Multiplexed Scanned Temporal Focusing (MuST)

 描述(由申请人提供)：我们建议设计和制造一种设备，该设备将突破现有技术的边界，以高速和单细胞分辨率对大规模神经元网络的活动进行成像。这将通过使用多路扫描时间聚焦(MASH)策略与具有优化脉冲特性的激光系统相结合来实现。我们的方法将能够以多赫兹的时间分辨率对空前大的V-FOV(20赫兹时为500x500x500微米，或3赫兹时为1x1x0.7毫米)进行无偏钙成像。这样的成像系统有可能给神经科学带来革命性的变化，首次提供了几乎同时监测数万个神经元网络活动动态的能力。在哺乳动物的大脑皮层中，这种能力提供了洞察信息处理的计算原理的机会，因为它将允许捕获跨大脑皮层的网络活动的动态关联。我们的技术将通过迭代和用户主导的方式与哥伦比亚大学的合作者一起开发，在那里神经科学界将可以访问我们的技术。将通过提供一个专门的网站进一步促进这项技术的传播，该网站包括开放源码以及详细的材料清单和设计和操作说明。

 DESCRIPTION (provided by applicant): We propose to design and build a device that will push the boundaries of existing technology for imaging the activity of large-scale neuronal networks at high speed and single cell resolution. This will be done by using a Multiplexed Scanned Temporal Focusing (MuST) strategy in combination with laser systems with optimized pulse characteristics. Our approach will enable unbiased calcium imaging of unprecedentedly large V-FOVs (500x500x500um at 20Hz, or 1x1x0.7mm at 3Hz) at multi-hertz time resolution. Such an imaging system has the potential to revolutionize neuroscience, providing, for the first time, the ability to monitor the dynamics of network activity of tens of thousands of neurons near-simultaneously. In the mammalian cortex this capability provides the opportunity to gain insights into the computational principles for information processing as it will allow capturing an correlating the dynamics of the network activity across cortical layers. Our technology will be developed through an iterative and in a user-led fashion with our collaborator at Columbia University where it will be made accessible to the neuroscience community. The dissemination of this technique will be further facilitated by providing a dedicated website including open source code and detailed list of materials and instructions for its design and operation.