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

该奖项是对单光子敏感共聚焦显微镜的开发，能够以视频速率进行真正的4D(四维：x，y，z，t)实时成像。它基于图像增强型CMOS传感器(ICMOS)和高速共焦扫描仪，旨在满足以下规格：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(电子倍增电荷耦合器件)的灵敏度相同，但帧速率快100倍。2)对单个x-y焦平面进行高速(1ms/帧)共焦扫描。此外，能够在30毫秒内进行高达100微米的深度(Z)扫描，从而以视频帧速率拍摄“真正的4D电影”。3)高速(10ms/帧)荧光寿命成像显微镜，单X-Y焦平面寿命分辨率为100PSEC。这与传统的扫描共聚焦显微镜的寿命分辨率相似，但帧速率快100倍。它将在视频帧速率下实现真正的4D电影。4)对单个x-y焦平面进行真光谱分析的视频率(~30ms/帧)胶片。这种新的显微镜可能会彻底改变在所有生物系统中显示毫秒级时间尺度现象的方式，从单分子、单细胞和神经网络(如大脑)到动物组织的活体成像。除了这种新型显微镜的科学益处外，该奖项还将有助于对生物、化学、物理和工程前沿的研究生和本科生进行多学科教育。

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.