MRI: Development of a High-Speed Confocal Microscope for 4D Live-Cell Imaging

MRI：开发用于 4D 活细胞成像的高速共焦显微镜

• 批准号: 0723204
• 负责人: Katsushi Arisaka
• 金额: $ 64.3万
• 依托单位: University of California-Los Angeles
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0723204&HistoricalAwards=false
• 关键词: MRI; Development; Speed; Confocal; Microscope

This award is for the development of a single-photon-sensitive confocal microscope, capable of true 4D (four dimensional: x, y, z, t) live imaging at video rate. It is based on an Image Intensified CMOS sensor (ICMOS) and a high-speed confocal scanner, which are designed to meet the following specifications: 1) High-speed ( 1 ms/frame), mega-pixel imaging with single-photon sensitivity. This is the same sensitivity as an EMCCD (Electron Multiplying CCD) but at one hundred times faster frame rate. 2) High-speed ( 1 ms/frame) confocal scanning for a single x-y focal plane. In addition, capability to scan in depth (z) up to 100 microns in 30 msec, resulting in a ''true 4D movie'' at video frame rate. 3) High-speed ( 10 ms/frame) FLIM (Fluorescence Lifetime Imaging Microscope) with 100 psec lifetime resolution for a single x-y focal plane. This is a similar lifetime resolution as conventional scanning confocal microscopes but with a frame rate that is one hundred times faster. It will enable a ''true 4D FLIM movie'' at video frame rate. 4) Video-rate (~30 ms/frame) FLIM with true spectrum analysis for a single x-y focal plane. This new microscope may revolutionize the way millisecond time-scale phenomena are visualized in all biological systems, spanning from single molecules, single cells, and neural networks (such as the brain), to in vivo imaging of tissue in animals. In addition to the scientific benefit of this new microscope, this award will contribute to multi-disciplinary education of students, at both the graduate and undergraduate level, at the forefront of biology, chemistry, physics, and engineering.

该奖项用于开发单光子敏感共聚焦显微镜，能够以视频速率进行真正的4D（四维：x， y, z, t）实时成像。它基于图像增强CMOS传感器（ICMOS）和高速共聚焦扫描仪，其设计满足以下规格：1)高速（1毫秒/帧），具有单光子灵敏度的百万像素成像。这与EMCCD（电子倍增CCD）具有相同的灵敏度，但帧速率要快100倍。2)对单个x-y焦平面进行高速（1ms /帧）共聚焦扫描。此外，能够在30毫秒内扫描深度(z)高达100微米，从而以视频帧速率产生“真正的4D电影”。3)高速（10毫秒/帧）荧光寿命成像显微镜（FLIM），单个x-y焦平面寿命分辨率为100秒。这与传统扫描共聚焦显微镜的寿命分辨率相似，但帧速率快了100倍。它将以视频帧率实现“真正的4D电影”。4)视频速率（~ 30ms /帧）具有单个x-y焦平面的真实光谱分析的FLIM。这种新型显微镜可能会彻底改变所有生物系统中毫秒时间尺度现象的可视化方式，从单个分子、单个细胞和神经网络（如大脑）到动物组织的体内成像。除了这种新型显微镜的科学效益外，该奖项还将为研究生和本科生在生物学、化学、物理学和工程学前沿的多学科教育做出贡献。