DEVELOPMENT OF NEW AUTOMATED POLARIZED LIGHT MICROSCOPE

我们正在开发一种新的、自动化的偏振光显微镜(POL- 范围“)，这极大地增强了 传统的Poll-Scope。新系统目前的能力包括： 样品延迟性的测量，而不考虑取向和 在全空间分辨率下，同时显示整个视场 光学显微镜的灵敏度为0。L nm延迟效应 1度的震级和角度分辨率。在以下位置完成图像记录 单焦点级别在0.43秒内获得，记录良好 在薄的光学部分内的结构和分子组织 样本。 我们建议加强现有文书的功能，包括 至少三个重要方面：数据采集速度(运行时间 等于约10毫秒)、灵敏度(0.02 nm延迟)和空间 分辨率(所有3个维度均为0.2微摩尔)。增强的空间 延迟测量的分辨率将通过 三维光学切片反褶积算法的研究进展 偏振光图像。所有这些改进，其他类型的 光学显微镜也将受益，将大大拓宽 新极镜的应用，特别是在3-甲基-4-羟基苯乙酸乙酯的研究中 活细胞中的维度建筑动力学。具体来说，我们 建议使用改进的系统测量：(A)微管发育迟缓 (体外和体内)作为微管动力学和 与其他蛋白质结合(MAP‘s)；(B)微管和其他蛋白质 延迟作为波长的函数来探索潜在的 作为化学成分指示剂的延迟分散；(C)3-D 有丝分裂纺锤体的结构及其与位置和位置的关系 纺锤体极和染色体的运动；(D)染色体和DNA三级 (去凝聚)精子的结构；和(E)空间和时间 病毒液晶中的双折射涨落决定了 具有良好特征的模型系统的动力学和粘弹性性质 水悬浮液中分子聚集体的有序阵列。我们就是这样 提议继续并加强已经在进行的合作努力 在大多数指定的应用领域。

We are developing a new and automated polarized light microscope ("pol- scope") which dramatically enhances the unique capabilities of the traditional pol-scope. Current capabilities of the new system include: Measurement of specimen retardance, regardless of orientation and concurrently for the whole field of view, at the full spatial resolution of the light microscope, with a sensitivity of 0. l nm retardance magnitude and angular resolution of 1 degree. Complete image records at a single focus level are obtained in 0.43 seconds documenting fine structural and molecular organization within a thin optical section of the specimen. We propose to enhance the capabilities of the existing instrument in at least three significant areas: speed of data acquisition (elapsed time equals approximately 10 ms), sensitivity (0.02 nm retardance), and spatial resolution (0.2 micromoles in all 3 dimensions). Enhanced spatial resolution of retardance measurements will be achieved through the development of 3-D deconvolution algorithms for optical sections of polarized light images. All these improvements, from which other types of light microscopy will benefit as well, will substantially broaden the applications of the new pol-scope, especially for the study of 3- dimensional architectural dynamics in living cells. Specifically, we propose to measure with the improved system: (a) microtubule retardance (in vitro and in vivo) as a function of microtubule dynamics and association with other proteins (MAP's); (b)microtubule and other protein retardances as a function of wavelength to explore the potential of retardance dispersion as indicator for chemical composition; (c) the 3-D architecture of the mitotic spindle and its relationship to position and movement of spindle poles and chromosomes; (d) chromosome and DNA tertiary structure of (decondensing) sperm; and (e) spatial and temporal birefringence fluctuations in virus liquid crystals determining the dynamic and viscoelastic properties of well-characterized model systems of ordered arrays of molecular aggregates in aqueous suspension. We are thus proposing to continue and intensify collaborative efforts already underway in most of the named application areas.