Regulation of Epidermal Junctions in C. elegans

描述（由申请人提供）： 我们的实验室开发了基于Java的，可免费分发的开源软件工具，用于捕获，整理，分析和传播从高分辨率Nomarski（DIG），多光子，激光扫描共聚焦和旋转盘共聚焦拍摄实验中获得的四维（4D）数据集，利用基于Java的NIH赞助的应用程序ImageJ的强大的开源架构。我们访问采集硬件的关键变化以及我们软件所依赖的底层Java API的变化提供了大幅升级这些软件应用程序的机会，这反过来又将使生物医学研究人员能够以具有成本效益的方式轻松创建和交换4D数据集。我们在PA-02-141计划公告中提出了以下补充子目标：补充子目标1：完成现有的ImageJ插件，用于回放和生产基于QuickTime的4D数据集。我们有QT 4D Player的测试版，这是一个用于查看和操作4D数据集的ImageJ插件，还有QT 4D Writer，这是一个压缩4D数据集以供QT 4D Player使用的应用程序。这两个独特而强大的跨平台应用程序，在我们的实验室不断使用了一年多，将被抛光和传播。补充Subaim 2：完成ImageJ插件，用于多种4D文件格式的数据导入和导入数据集的后续3D重建：我们有一个转换套件的工作alpha版本，可以生成4D数据的旋转3D重建，然后压缩它们以进行回放;我们将完成这个套件。补充Subaim 3：使用IIDC Firewire相机标准调整现有Java本地接口（JNI）代码以进行4D采集。我们已经制作了一个最初为模拟图像采集器编写的4d采集包的测试版。我们将使用为Mac和Windows操作系统开发的现有JNI库，调整此套件以支持Prior聚焦电机和火线CCD相机。这些新增功能将允许相同的设备获取4D荧光（旋转磁盘）或Nomarski数据集。这些4D工具的开发将使我们的实验室能够继续追求其目标，即在活体C中对荧光标记分子进行实时4D成像。线虫胚胎此外，这个免费提供的开源套件将允许研究人员和教育工作者使用易于扩展的、平台无关的Java技术轻松、廉价地访问4D分析和获取功能。

DESCRIPTION (provided by applicant): Our laboratory has developed Java-based, freely distributable, open-source software tools for capturing, collating, analyzing, and disseminating four-dimensional (4d) datasets acquired from high-resolution Nomarski (DIG), multiphoton, laser scanning confocal, and spinning disk confocal filming experiments, leveraging the powerful, open-source architecture of the Java-based NIH-sponsored application, ImageJ. Keys changes in our access to essential pieces of acquisition hardware and changes in the underlying Java APIs upon which our software relies provide an opportunity to substantially upgrade these software applications, which in turn will allow biomedical investigators to create and exchange 4d datasets easily in a cost-effective manner. We propose the following supplementary subaims, under program announcement PA-02-141: Supplementary Subaim 1: Completion of existing ImageJ plugins for playback and production QuickTime-based, 4d datasets. We have working beta versions of QT4D Player, an ImageJ plugin for viewing and manipulating 4d datasets, and QT4D Writer, an application that compresses 4d datasets for use in QT4D Player. These two unique and powerful cross-platform applications, in constant use in our laboratory for over a year, will be polished and disseminated. Supplementary Subaim 2: Completion of ImageJ plugins for data import of multiple 4d file formats and subsequent 3d reconstruction of imported datasets: We have a working alpha version of a conversion suite that can produce rotated, 3d reconstructions of 4d data and subsequently compress them for playback; we will complete this suite. Supplementary Subaim 3: Adaptation of existing Java Native Interface (JNI) code for 4d acquisition using the IIDC Firewire camera standard. We have produced a beta version of a 4d acquisition package originally written for analog frame grabbers. We will adapt this suite to support Prior focus motors and Fire Wire CCD cameras, using existing JNI libraries developed for the Mac and Windows OS. These additions will allow the same apparatus to acquire either 4d fluorescence (spinning disk) or Nomarski datasets. The development of these 4d tools will allow our laboratory to continue to pursue its goals of live, 4d imaging of fluorescently tagged molecules in living C. elegans embryos. Moreover, this freely available, open-source suite will allow investigators and educators easy and inexpensive access to 4D analysis and acquisition capabilities using easily extensible, platform-independent Java technologies.