Novel sub-cellular chemical and mechanical nanoimaging

描述(由申请人提供)：我们建议开发一个成像平台，以前所未有的分辨率绘制细胞和亚细胞结构的化学特性和机械行为。目前，透射式电子显微镜和荧光显微镜技术都在经历一场化学选择性和空间分辨率的革命。然而，无论是TEMS都不能成像溶液中的功能生物系统，荧光显微镜也不能提供原子尺度的分辨率。我们建议的成像平台使用一种完全不同的方法来提供溶液环境中埃斯特罗姆尺度的化学信息。我们开发了一种在生物小分子的能量景观中编码化学信息的方案，并开发了一种纳米机械设备来快速解码这些信息。化学信息可以通过基因操作被编码到靶分子中，这将导致绿色荧光蛋白技术的机械模拟。我们的原理实验证明，在溶液环境中，可以获得亚纳米分辨率的生物分子的化学特定的多色图像。我们建议将这一概念发展成一个成像平台，可以针对活细胞和分离的复合体中的各种蛋白质和其他生物分子。新成像平台的能力将超越化学识别，并允许探测生物分子之间的相互作用，生物分子共同决定细胞的机械行为。对细胞生物状态的准确描述不仅仅包括其生化成分及其空间排列，因为细胞的机械行为会影响正常和疾病的细胞过程。新的成像平台将以前所未有的空间和时间分辨率提供有关局部压缩-拉伸和弹性-粘弹性特性的信息，从而确定细胞的机械状态。我们将进一步将新的纳米机械成像平台与包括超分辨方法在内的荧光显微镜相结合，剖析观察到的动态力学特征的分子基础。一旦开发出来，我们将把这项技术应用于细胞力学、黏附、突触可塑性和大分子复合体的结构确定等问题。该项目开发的技术和仪器将在广泛的生物医学领域和技术中产生广泛的影响。

DESCRIPTION (provided by applicant): We propose to develop an imaging platform to map chemical identities and mechanical behaviors across cells and sub-cellular structures with unprecedented resolution. Currently, both transmission electron microscopy (TEM) and fluorescence microscopy technologies are going through a revolution, one in chemical selectivity and the other in spatial resolution. However, neither TEMs can image functional biological systems in solution nor fluorescence microscopy can provide atomic scale resolution. Our proposed imaging platform uses a radically different approach to provide chemical information on the Angstrom scale in solution environment. We developed a scheme to encode chemical information in energy landscapes of small biomolecules and a nanomechanical device to decode this information rapidly. The chemical information can be encoded into target molecules through genetic manipulations, which will lead to the mechanical analogue of the green fluorescent protein technology. Our proof of principle experiments have demonstrated that chemically-specific multicolor images of biomolecules can be obtained with sub-nanometer resolution in a solution environment. We propose to develop this concept into an imaging platform that can target a wide variety of proteins and other biomolecules across living cells and in isolated complexes. The capabilities of the new imaging platform will go beyond chemical identification and allow probing interactions among biomolecules, which collectively determine the mechanical behavior of cells. An accurate description of the biological state of a cell has to include more than its biochemical composition and their spatial arrangements, because mechanical behaviors of cells influence normal and diseased cellular processes. The new imaging platform will allow determining the mechanical state of cells by providing information on local compression-tension and elastic-viscoelastic characteristics at unprecedented spatial and temporal resolution. We will further combine the new nanomechanical imaging platform with fluorescence microscopy including super-resolution methods to dissect the molecular basis of the observed dynamic mechanical characteristics. Once developed, we will apply this technology to problems in cell mechanics, adhesion, synaptic plasticity and structure determination of macromolecular complexes. The technology and instrumentation developed in this project will have a broad impact in a wide range of biomedical fields and technologies.