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，透射电子显微镜（TEM）和荧光显微镜技术都在经历一场革命，一个是化学选择性，另一个是空间分辨率。然而，tem不能在溶液中成像功能性生物系统，荧光显微镜也不能提供原子尺度的分辨率。我们提出的成像平台使用了一种完全不同的方法来提供溶液环境中埃尺度的化学信息。我们开发了一种方案来编码小生物分子能量景观中的化学信息，并开发了一种纳米机械装置来快速解码这些信息。化学信息可以通过基因操作编码到目标分子中，这将导致绿色荧光蛋白技术的机械模拟。我们的原理证明实验表明，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异性多色图像。我们建议将这一概念发展成一种成像平台，可以针对活细胞和孤立复合物中的各种蛋白质和其他生物分子。新成像平台的功能将超越化学鉴定，并允许探测生物分子之间的相互作用，这些相互作用共同决定了细胞的机械行为。对细胞生物状态的准确描述必须包括其生化组成及其空间排列，因为细胞的机械行为影响正常和病变的细胞过程。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-张力和弹性-粘弹性特性的信息，从而确定细胞的机械状态。我们将进一步结合新的纳米力学成像平台与荧光显微镜，包括超分辨率方法，剖析观察到的动态力学特性的分子基础。一旦开发成功，我们将把这项技术应用于细胞力学、粘附、突触可塑性和大分子复合物结构确定等问题。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.