Imaging Polarimeter for Simultaneous Determination of Luminescence Polarization

描述(申请人提供)：圆二色谱(CD)是分析药物及其与生物聚合物结合的重要分析工具。它是基于手性分子，如许多小分子药物，以及蛋白质和核酸对左右螺旋光的差异吸收。不幸的是，CD和与发光相关的现象--发光--荧光检测的圆二色(FDCD)和圆偏振发射(CPE)--不适用于高通量的发现平台，在这种平台中，药物根据各种合成条件进行筛选，或者在其与各种生物活性受体的相互作用中进行筛选。这是因为光学分析中常用的塑料多孔板具有残余应变和相关的折射各向异性，这破坏了左右圆偏振光传输的比较。在这里，我们的目标是建立一种仪器，即使在多孔板上也可以用CD分析来筛选药物。该仪器需要与井板相适应的垂直设计。它还依赖于样品前后光的偏振态的一个以上的调制器，以及在分析在这种布置期间产生的复杂信号方面的必要经验。(通常CD光谱仪使用单个调制器)。该仪器将具有重要意义，因为CD、FDCD和CPE都是药物-生物分子相互作用的强大探针。因此，基于同时获取这些参数的高通量筛选分析将广泛影响生物医学领域。它将是强大的，因为同时收集各种不同但相关的物理现象将提高我们分析的准确性，并消除反筛查的需要，以进一步区分阳性结果。我们进一步设计了一种对这些参数进行成像的策略，即使它们的测量基于比成像相机更快的偏振调制器。这一策略的核心是频闪LED阵列，该阵列还用于计算推导必要的光学特性所需的傅里叶分析。通过合成化学文库，筛选现有化合物，或通过与特定靶点结合的有效性分析，越来越多的药物以连续的方式被发现。我们的仪器具有公共利益，可以加快从发现活性化合物到发现与各种疾病相关的活性药物的过程。我们将通过检测溶液和生物体中的DNA和染色质，以及它们在已知影响核酸构象变化的试剂存在时的反应，来测试我们所谓的米勒矩阵荧光仪的有效性。

DESCRIPTION (provided by applicant): Circular dichroism (CD) is an important analytical tool for the analysis of drugs and their binding to biopolymers. It is based on the differential absorption of left and right helical light by chiral molecules such as many small molecule drugs, as well as proteins and nucleic acids for examples. Unfortunately, CD, and related phenomena that are related to luminescence -- the emission of light -- fluorescence detected circular dichroism (FDCD) and circularly polarized emission (CPE) of light, are not amenable to high-throughput discovery platforms in which a drug is screened against a variety of conditions of synthesis, or in its interactions with a variety of biological active receptors. This is because th plastic multi-well plates commonly used in optical analysis have residual strain and associated refraction anisotropy that corrupts comparisons of transmission of left and right circularly polarized light. Here, we aim to build an instrument that can be used for screening drugs with CD analysis even in multi-well plates. The instrument requires a vertical design adapted to well plates. It also relies on more than one modulator of the polarization state of light before and after the sample and the necessary experience with analyzing the complex signal that is produced during such an arrangement. (Typically CD spectrometers use a single modulator). The instrument will be significant because CD, FDCD, and CPE are all powerful probes of drug-biomolecule interactions. Thus, high-throughput screening assays based on the simultaneous acquisition of these parameters will broadly impact the biomedical field. It will be powerful because the simultaneous collections of a variety of distinct but related physical phenomena will increase the accuracy of our assays and obviate the need for counter-screening to further differentiate positive hits. We further devise a strategy for imaging each of these parameters, even though their measurement is based on polarization modulators that are faster than imaging cameras. At the heart of this strategy is a stroboscopic LED array that also serves to compute the Fourier analysis necessary for deriving the requisite optical properties. Drugs are increasing discovered in serial fashion through synthesis of via chemical libraries, screening of existing compounds, or via assays of efficacy with respect to the binding to a particular target. Our instrument with be of public benefit in speeding up the pipeline from discovery of active compounds to active pharmaceuticals associated with all manner of disease and illness. We will test the efficacy of our so-called Mueller matrix fluorimeter by examining DNA and chromatin in solution, and in organisms, and their responses in the presence of agents known to affect conformational changes in nucleic acids.