Novel sub-cellular chemical and mechanical nanoimaging

描述（由申请人提供）：我们建议开发一种成像平台，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，一场是在化学选择性方面，另一场是在空间分辨率方面。然而，TEM既不能在溶液中成像功能生物系统，也不能提供原子尺度的分辨率。我们提出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设备来快速解码这些信息。化学信息可以通过基因操作编码到靶分子中，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，可以在溶液环境中以亚纳米分辨率获得生物分子的化学特异性荧光图像。我们建议将这一概念发展成一个成像平台，可以靶向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，这些生物分子共同决定细胞的机械行为。对细胞生物学状态的准确描述必须包括其生物化学组成及其空间排列，因为细胞的机械行为影响正常和患病的细胞过程。新的成像平台将允许通过以前所未有的空间和时间分辨率提供关于局部压缩-张力和弹性-粘弹性特征的信息来确定细胞的机械状态。我们将进一步联合收割机结合新的纳米机械成像平台与荧光显微镜，包括超分辨率的方法，解剖所观察到的动态力学特性的分子基础。一旦开发出来，我们将把这项技术应用于细胞力学、粘附、突触可塑性和大分子复合物的结构确定等问题。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.