Ultrafast Bioimaging

项目概述：PI建议开发一个超快生物成像程序，该程序可以打开一个新的领域 并导致了一系列基本的科学发现。空间和时间，两个关键的物理 尺寸，构成了现代计量学的基础。在生物成像领域，2014年诺贝尔奖 在化学方面，在提高显微成像的空间分辨率方面取得了惊人的进展， 这就产生了一大批可以突破光的衍射极限的纳米工具。尽管同样 重要的是，追求高时间分辨率只是最近才引起注意，这要归功于 几种使能技术。开发这些超快成像仪的动机来自于 当代生物学的景观转移，从形态学探索和表型探索 生物体寻求定量的见解，在分子水平上的潜在机制。瞬态 分子事件发生在配体结合所需的几十到几百微秒的时间尺度上， 到几十飞秒的分子振动时间。因此，超快成像对于观察至关重要 以及这种动态事件的特征。 因此，在微观尺度上的大多数超快现象都是使用非成像技术探测的。 方法.然而，由于大多数瞬时分子事件是分子级联的结果， 由于这些互动不是孤立发生的，缺乏图像限制了分析的范围。另 一方面，尽管能够捕获二维图像，但基于电子照相机的传统照相机 诸如CCD和CMOS之类的图像传感器不能在期望的成像下提供高帧速率 由于电子带宽限制（数据传输、数字化和写入）， 为了解决这个根本问题，我们的策略是将压缩感知的范式引入到 高速光学成像而不是测量事件数据立方体的每个时空体素，我们将 利用生物场景的可压缩性，从而更有效地利用相机的带宽- 图像数据在被数字化并传送到主机之前被压缩。此功能将 使我们的方法特别有利于记录高速图像数据，否则将 如果在奈奎斯特采样下测量，则需要巨大的相机带宽和硬件资源。基于 这一策略，我们将探索超快生物成像的帧速率从几MHz到十太赫兹，范围是 对于理解生物分子行为至关重要，但目前无法通过传统的高速 相机由此产生的研究计划将最终导致新一代超快生物成像仪， 在最先进的方法上取得革命性的进步。

Project Summary: The PI proposes to develop an ultrafast bioimaging program which could open a new area of investigation and lead to a series of fundamental scientific discoveries. Space and time, two key physical dimensions, constitute the basis of modern metrology. In bio-imaging, as recognized by the 2014 Nobel Prize in chemistry, there have been breathtaking advances in improving the spatial resolution of microscopic imaging, resulting in an impressive arsenal of nanoscopy tools that can break the diffraction limit of light. Despite equally important, the pursuit of a high-temporal resolution has only recently caught attention thanks to the emergence of several enabling technologies. The motivation to develop these ultrafast imagers originates from the landscape shift of the contemporary biology from morphological explorations and phenotypic probing of organisms to seeking quantitative insights into underlying mechanisms at molecular levels. The transient molecular events occur at a timescale varying from tens and hundreds of microseconds that ligands take to bind, to tens of femtoseconds that molecules take to vibrate. Ultrafast imaging, therefore, is essential for observation and characterization of such dynamic events. Heretofore, most ultrafast phenomena at microscopic scales were probed using non-imaging-based methods. However, since most transient molecular events are a consequence of a cascade of molecular interactions, rather than occurring in isolation, the lack of images limits the scope of the analysis. On the other hand, despite the capability of capturing two-dimensional images, conventional cameras based on electronic image sensors, such as CCD and CMOS, fall short in providing a high frame rate under desirable imaging conditions due to electronic bandwidth limitations (data transfer, digitalization, and writing). To solve this fundamental problem, our strategy is to introduce the paradigm of compressed sensing into high-speed optical imaging. Rather than measuring each spatiotemporal voxel of an event datacube, we will leverage the compressibility of biological scenes and thereby utilizes the camera’s bandwidth more efficiently— the image data is compressed before being digitalized and transferred to the host computer. This feature will make our approaches especially advantageous for recording high-speed image data, which otherwise would require tremendous camera bandwidth and hardware resources if measured under Nyquist sampling. Based on this strategy, we will explore ultrafast bioimaging at a frame rate from a few MHz to ten THz, a range which is essential for understanding the biomolecular behaviors but currently inaccessible by conventional high-speed cameras. The resultant research program will ultimately lead to a new generation of ultrafast bioimagers and make transformative advancements to the state-of-the-art methods.