SCAPE microscopy for high-speed in-vivo volumetric microscopy in behaving organisms

SCAPE 显微镜用于行为生物体的高速体内体积显微镜

• 批准号: 9328178
• 负责人: Elizabeth M. C. Hillman
• 金额: $ 50.48万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-30 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9328178
• 关键词: 3-Dimensional; Address; Adoption; Adult; Apical; Area; Behavior; Behavioral; Behavioral Assay; Biomedical Research; Brain; Brain imaging; Caenorhabditis elegans; Cells; Collaborations; Complex; Computer software; Data; Dendrites; Development; Drosophila genus; Drosophila melanogaster; Event; Genetic; Geometry; Hybrids; Image; Imaging Techniques; Individual; Institutes; Laboratories; Larva; Laser Scanning Microscopy; Lasers; Legal patent; Light; Lighting; Microscopy; Mind; Modeling; Motion; Motor; Mus; Nature; Nervous system structure; Neurobiology; Neurons; Neurosciences Research; Noise; Optics; Organism; Pattern; Penetration; Performance; Photons; Play; Publishing; Research Personnel; Resolution; Rewards; Rodent; Role; Sampling; Scanning; Signal Transduction; Somatosensory Cortex; Speed; Structure; System; Task Performances; Techniques; Testing; Three-Dimensional Imaging; Time; Tissues; Translating; Translations; Walking; Work; Zebrafish; awake; base; behavioral response; brain behavior; cellular imaging; cost; fly; genetic manipulation; high risk; imaging approach; imaging platform; improved; in vivo; information processing; interest; lens; neural circuit; neuroimaging; novel strategies; optogenetics; photonics; prototype; public health relevance; relating to nervous system; somatosensory; spatiotemporal; tool; two-photon; user-friendly; whole body imaging

 DESCRIPTION (provided by applicant): Despite the growing availability of optical markers of neuronal activity, as well as genetic tools for optical manipulation, current optical microscopy techniques for imaging the intact brain at cellular resolution have approached their limits, particularly in terms of 3D volumetric imaging speeds. The brain and nervous system is inherently 3D, with cortical layers playing specific roles in information processing. Small organisms such as Drosophila melanogaster (fruit fly), Danio rerio (zebrafish) and Caenorhabditis elegans, have become valuable platforms for neuroscience research and genetic manipulation, and offer the chance to capture the entire nervous system of a complete, behaving organism. However, for both rodent brain and small organism microscopy, current techniques are limited to slow volumetric imaging rates, or single-plane acquisition. We recently developed a transformative new approach to high speed 3D microscopy called Swept, Confocally-Aligned Planar Excitation (SCAPE) microscopy. SCAPE was conceived as a way to dramatically improve volumetric imaging speeds, while maintaining a simple optical layout and image acquisition geometry. SCAPE is a hybrid between light-sheet microscopy and laser scanning confocal which overcomes the major speed barriers of both techniques. Recently published in Nature Photonics, SCAPE can image at volume rates 10-100 x faster than laser scanning microscopy or fast light-sheet imaging. We have demonstrated imaging of cellular-level structure and function in both the awake, behaving rodent brain and freely moving Drosophila melanogaster larvae at 10-20 volumes per second (VPS) over large fields of view. A further feature of SCAPE is its simple, single, stationary objective, permitting 3D imaging with no motion at the sample, making it well suited for integration with pattered optogenetic manipulation of cells during high-speed 3D imaging. Having achieved `proof of concept' we now wish to develop SCAPE into a tool for routine use by neuroscientists working in both small organisms, for in-toto imaging of cellular activity and behavior, and in awake, behaving mouse brain. The former will be achieved through development and translation of an improved beta prototypes `1P-SCAPE' system, with development of user friendly acquisition software, data handling and analysis platforms, and ultimately its deployment and support for use in studies of somatosensory integration in adult and larval Drosophila. For mouse brain imaging, we propose to test the limits of SCAPE by exploring two- photon implementation (2P-SCAPE), which will afford deeper penetration imaging into scattering tissues such as the rodent brain.

 描述（由申请人提供）：尽管神经元活动的光学标记以及用于光学操作的遗传工具越来越多，但当前以细胞分辨率对完整大脑进行成像的光学显微镜技术已接近其极限，特别是在 3D 体积方面大脑和神经系统本质上是 3D 的，皮质层在信息处理中发挥着特定的作用，例如果蝇、斑马鱼和斑马鱼。秀丽隐杆线虫已成为神经科学研究和基因操作的宝贵平台，并提供了捕获完整的、有行为的生物体的整个神经系统的机会。然而，对于啮齿类动物的大脑和小型生物体的显微镜检查，当前的技术仅限于慢速体积成像。我们最近开发了一种革命性的高速 3D 显微镜新方法，称为扫描共焦平面激发 (SCAPE) 显微镜。 SCAPE 被认为是一种显着提高体积成像速度的方法，同时保持简单的光学布局和图像采集几何形状。SCAPE 是光片显微镜和激光扫描共焦的混合体，克服了这两种技术的主要速度障碍。 Nature Photonics 的 SCAPE 成像速度比激光扫描显微镜或快速光片成像快 10-100 倍。我们已经证明了清醒、行为啮齿动物的细胞水平结构和功能的成像。 SCAPE 的另一个特点是其简单、单一、固定的物镜，可以在大视野范围内以每秒 10-20 体积 (VPS) 的速度对大脑和自由移动的果蝇幼虫进行 3D 成像。 样本的运动，使其非常适合在高速 3D 成像过程中与细胞的图案光遗传学操作集成。在实现“概念验证”后，我们现在希望将 SCAPE 开发成一种供研究小型生物体的神经科学家日常使用的工具。 ，用于细胞活动和行为的整体成像，以及清醒、行为小鼠大脑的成像。前者将通过开发和翻译改进的测试原型“1P-SCAPE”系统来实现，并开发用户友好的采集功能。软件、数据处理和分析平台，以及最终其在成年和幼虫果蝇体感整合研究中的部署和支持。对于小鼠大脑成像，我们建议通过探索双光子实现（2P-SCAPE）来测试 SCAPE 的极限。 ），这将为啮齿动物大脑等散射组织提供更深入的穿透成像。

项目成果

Neuromuscular Basis of Drosophila larval rolling escape behavior.

果蝇幼虫滚动逃逸行为的神经肌肉基础。

• 发表时间: 2023-08-24
• 作者: Cooney, Patricia C;Huang, Yuhan;Li, Wenze;Perera, Dulanjana M;Hormigo, Richard;Tabachnik, Tanya;Godage, Isuru;Hillman, Elizabeth M C;Grueber, Wesley B;Zarin, Aref A
• 通讯作者: Zarin, Aref A

Whole-Volume Clustering of Time Series Data from Zebrafish Brain Calcium Images via Mixture Modeling.

通过混合建模对斑马鱼脑钙图像的时间序列数据进行全体积聚类。

• 发表时间: 2018-02
• 作者: Nguyen, Hien D;Ullmann, Jeremy F P;McLachlan, Geoffrey J;Voleti, Venkatakaushik;Li, Wenze;Hillman, Elizabeth M C;Reutens, David C;Janke, Andrew L
• 通讯作者: Janke, Andrew L

The spatial and temporal structure of neural activity across the fly brain.

果蝇大脑神经活动的空间和时间结构。

• 发表时间: 2023-09-11
• 影响因子: 16.6
• 作者: Schaffer, Evan S;Mishra, Neeli;Whiteway, Matthew R;Li, Wenze;Vancura, Michelle B;Freedman, Jason;Patel, Kripa B;Voleti, Venkatakaushik;Paninski, Liam;Hillman, Elizabeth M C;Abbott, L F;Axel, Richard
• 通讯作者: Axel, Richard

Odorant Receptor Inhibition Is Fundamental to Odor Encoding.

气味受体抑制是气味编码的基础。

• 发表时间: 2020-07-06
• 作者: Pfister, Patrick;Smith, Benjamin C;Evans, Barry J;Brann, Jessica H;Trimmer, Casey;Sheikh, Mushhood;Arroyave, Randy;Reddy, Gautam;Jeong, Hyo;Raps, Daniel A;Peterlin, Zita;Vergassola, Massimo;Rogers, Matthew E
• 通讯作者: Rogers, Matthew E

The mesencephalic locomotor region recruits V2a reticulospinal neurons to drive forward locomotion in larval zebrafish.

中脑运动区招募 V2a 网状脊髓神经元来驱动斑马鱼幼虫的向前运动。

• 发表时间: 2023-10
• 影响因子: 25
• 作者: Carbo;Lapoix, Mathilde;Jia, Xinyu;Thouvenin, Olivier;Pascucci, Marco;Auclair, François;Quan, Feng B;Albadri, Shahad;Aguda, Vernie;Farouj, Younes;Hillman, Elizabeth M C;Portugues, Ruben;Del Bene, Filippo;Thiele, Tod R;Dubuc, Réje
• 通讯作者: Dubuc, Réje