DEVELOPMENT OF NEW AUTOMATED POLARIZED LIGHT MICROSCOPE

我们正在开发一种新的自动偏振光显微镜（“pol-light”）。 范围”），这极大地增强了 传统的pol-scope新系统目前的功能包括： 测量试样延迟，无论取向如何， 在全空间分辨率下，同时针对整个视场 光学显微镜，灵敏度为0。1 nm延迟 幅度和1度的角分辨率。在一个完整的图像记录 0.43秒内获得单焦点级别，记录良好 结构和分子组织内的薄光学部分的 试样 我们建议加强现有文书的能力， 至少有三个重要方面：数据采集速度（运行时间 等于约10 ms）、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 光学断面三维反卷积算法的发展 偏振光图像所有这些改进，从其他类型的 光学显微镜也将受益，将大大拓宽 新的pol-scope的应用，特别是对3- 活细胞中的三维结构动力学。我们特别 建议用改进的系统测量：（a）微管阻滞 (in体外和体内）作为微管动力学的函数， 与其它蛋白质的结合（MAP）;（B）微管和其它蛋白质 延迟作为波长的函数，以探索 延迟分散作为化学成分的指标;（c）三维 有丝分裂纺锤体的结构及其与位置和 纺锤体极和染色体的运动;（d）染色体和DNA三级结构 （解致密）精子的结构;以及（e）空间和时间 病毒液晶中的双折射波动决定了 动态和粘弹性特性的良好表征的模型系统， 有序阵列的分子聚集体在水悬浮液。因此我们 建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.