Numerical Instrumentation for Shape and Shape Change Analysis via Three-Dimensional Microscopy

研究人员将把研究重点放在全面开发微观系统的自动化仪器功能上，该系统提供了量化微观尺度生物标本的三维形状参数和形状参数随时间变化的潜力。他们计划继续研究互补成像方式的多样性，包括共聚焦显微镜、立体显微镜和传统光学切片显微镜。研究项目将包括通过各种方法获取微观数据。所获得的图像将根据图像形成的物理原理进行适当的滤波方案，以锐化和提高单个显微照片的图像清晰度，并去除对局部标本结构的虚假信息。滤光技术在其发展过程中需要大量的分析研究，将针对所使用的每个光学系统。最后，聚合的本地光学数据将通过复杂的数值仪器技术进行组装和分析，以重建给定成像标本的三维结构，并量化其尺寸和形状特征，因为它随着时间的推移而演变。开发的方法将在广泛的标本类型上实施和评估，包括染色和未染色的单细胞和多细胞受试者。几种不同的组织制剂将用于研究标本形态随时间在各种生物过程中的时间演变。

The investigators will focus their research on fully developing the automated instrumentation capabilities of microscopic systems that offer the potential for quantifying the 3-D shape parameters, and changes in shape parameters over time, of microscopic-scale biological specimens. They plan to continue their program of studying diversity of complementary imaging modalities, including confocal microscopy, stereo microscopy, and conventional optical sectioning microscopy. The program of study will encompass microscopic data acquisition via various methods. The images obtained will be subjected to appropriate filtering schemes base on the physical principles of image formation to sharpen and improve image clarity in individual micrographs and to remove information spurious to that of the local specimen structure. The filtering techniques, which will require considerable analytic study in their development, will be specific to each optical system used. Finally, the aggregate local optical data will be assembled and analyzed via sophisticated numerical instrumentation techniques to reconstruct the 3-D structure of a given imaged specimen and to quantify its size and shape characteristics as it evolves over time. The methods developed will be implemented and evaluated on a broad range of specimen types, including stained and unstained single and multiple celled subjects. Several different tissue preparations will be used to study the time evolution of specimen morphology over time during various biological processes.