High Speed, Multi-sensor Light Field Deconvolution Microscopy for Whole Brain Recording of Neuronal Activity

用于全脑记录神经元活动的高速、多传感器光场解卷积显微镜

• 批准号: 9222798
• 负责人: Edward S. Boyden
• 金额: $ 44.23万
• 依托单位: LEAFLABS, LLC
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-02-18 至 2019-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9222798
• 关键词: Algorithms; Alzheimer' s Disease; Anatomy; Animals; BRAIN initiative; Behavior; Biological; Brain; Brain Diseases; Calcium; Cognition; Computer software; Custom; Data; Data Set; Development; Engineering; Ensure; Epilepsy; Eye; Goals; Image; Laboratory Research; Learning; Light; Link; Memory; Mental Depression; Methods; Microscope; Microscopy; Monitor; Nervous system structure; Neurons; Neurosciences; Noise; Nuclear; Parkinson Disease; Polishes; Reporter; Resolution; Series; Signal Transduction; Speed; System; Technology; Time; Tissues; Video Recording; Willow; Work; Zebrafish; base; cost; data acquisition; density; design; fluorescence imaging; image reconstruction; imaging capabilities; imaging system; improved; millisecond; novel; programs; prototype; public health relevance; reconstruction; relating to nervous system; response; scale up; sensor; tool; two-photon; user-friendly; virtual reality; visual stimulus

 DESCRIPTION (provided by applicant): Recording all the neural activity in a 3D volume with millisecond timescale precision is a key goal of the BRAIN initiative. Recently, in a collaborative project with the Vaziri lab (IMP, Vienna), we adapted the strategy of lightfield microscopy for 3D volumetric imaging of fluorescent neural calcium responses (Prevedel 2014). This technology enables computational reconstruction of a 3D volume from an image by simultaneously capturing the angle of incident light rays in addition to their intensity. Imaging can occur as quickly as the fluorescent neural activity reporter allows (Chen 2013); we imaged the entire larval zebrafish brain at 20 Hz. However, the spatial resolution for lightfield microscopy is poo, resulting in low signal to noise ratio (SNR) as well as difficulty in automatically segmenting neural anatomy, which is key to linking neural activity to underlying circuitry. This spatial resolution limit is a fundamental issue with lightfield microscopy, since to gain 3­D imaging capability, one must sacrifice spatial resolution: there are only so many pixels on the camera. Accordingly, we here propose to perform the first whole brain recording of a larval zebrafish with single neuron resolution by increasing the total pixel count of our existing system by an order of magnitude whilst improving the SNR by leveraging a six fold increase in frame rate. Our existing lightfield imaging system (Prevedel 2014), and others (Levoy 2006, Cohen 2014), use an array of microlenses to effect the trade­off of spatial for axial resolution. An alternativ approach captures the lightfield using an array of cameras, without any microlenses. Our novel design combines both approaches at an unprecedented scale. Our scalable data acquisition system (Willow, see preliminary data) combined with cameras designed in­house limit the cost of our system to 1/10 of a traditional two­photon microscope. In this way we aim to develop a user­friendly system capable of imaging all of the neurons in a 3D volume, at speeds comparable to the natural timescales of neural activity, whilst keeping an eye towards polishing our system for marketability and widespread use.

 描述（由申请人提供）：以毫秒级时间尺度精度记录3D体积中的所有神经活动是BRAIN计划的关键目标。最近，在一个合作 在与Vaziri实验室（IMP，维也纳）合作的项目中，我们采用了光场显微镜的策略，用于荧光神经钙反应的3D体积成像（Prevedel 2014）。该技术能够通过同时捕获入射光线的角度以及它们的强度来从图像计算重建3D体积。成像可以在荧光神经活动报告器允许的情况下尽可能快地发生（Chen 2013）--我们以20 Hz对整个幼斑马鱼大脑进行成像。 然而，光场显微镜的空间分辨率很低，导致低信噪比（SNR）以及难以自动分割神经解剖结构，这是将神经活动与底层电路联系起来的关键。 这种空间分辨率限制是光场显微镜的一个基本问题，因为要获得3D成像能力，必须牺牲空间分辨率：相机上只有这么多像素。 因此，我们在这里建议通过将我们现有系统的总像素数增加一个数量级来执行具有单神经元分辨率的幼体斑马鱼的第一次全脑记录，同时通过利用帧速率的六倍增加来提高SNR。 我们现有的光场成像系统（Prevedel 2014）和其他系统（Levoy 2006，Cohen 2014）使用微透镜阵列来实现空间分辨率与轴向分辨率的权衡。另一种方法是使用相机阵列来捕获光场，而不使用任何微透镜。我们的新设计以前所未有的规模结合了这两种方法。我们的可扩展的数据采集系统（杨柳，见初步数据）与在Rumhouse设计的相机相结合，将我们的系统成本限制在传统双光子显微镜的1/10。通过这种方式，我们的目标是开发一个用户友好的系统，能够以与神经活动的自然时间尺度相当的速度对3D体积中的所有神经元进行成像，同时密切关注我们的系统的适销性和广泛使用。

项目成果

Scalable, Modular Three-Dimensional Silicon Microelectrode Assembly via Electroless Plating.

通过化学镀的可扩展、模块化三维硅微电极组件。

• DOI: 10.3390/mi9090436
• 发表时间: 2018
• 期刊: Micromachines
• 影响因子: 3.4
• 作者: Scholvin,Jörg;Zorzos,Anthony;Kinney,Justin;Bernstein,Jacob;Moore-Kochlacs,Caroline;Kopell,Nancy;Fonstad,Clifton;Boyden,EdwardS
• 通讯作者: Boyden,EdwardS