Imaging Polarimeter for Simultaneous Determination of Luminescence Polarization

描述（由申请人提供）：圆二色性（CD）是分析药物及其与生物聚合物结合的重要分析工具。它是基于手性分子对左右螺旋光的不同吸收，例如许多小分子药物，以及蛋白质和核酸。不幸的是，CD以及与发光有关的相关现象——光的发射——荧光检测到的圆二色性（FDCD）和光的圆偏振发射（CPE），不适合高通量发现平台，在这个平台上，药物是根据各种合成条件筛选的，或者是在它与各种生物活性受体的相互作用中。这是因为光学分析中常用的塑料多孔板存在残余应变和相关的折射各向异性，影响了左右圆偏振光透射的比较。在此，我们的目标是建立一种可以在多孔板中使用CD分析筛选药物的仪器。该仪器需要一个垂直的设计，以适应井板。它还依赖于样品前后光偏振状态的多个调制器，以及分析在这种安排过程中产生的复杂信号的必要经验。（典型的CD光谱仪使用单个调制器）。该仪器意义重大，因为CD， FDCD和CPE都是药物-生物分子相互作用的有力探针。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，并避免需要反筛选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，尽管它们的测量是基于比成像相机更快的偏振调制器。该策略的核心是频闪LED阵列，该阵列还用于计算导出所需光学特性所需的傅里叶分析。通过化学文库的合成，现有化合物的筛选，或通过与特定靶点结合的功效测定，药物的发现越来越多。我们的仪器在加速从发现与各种疾病和疾病相关的活性化合物到活性药物的管道方面具有公共利益。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下的反应，来测试我们所谓的穆勒矩阵荧光仪的功效。

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.