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. ? ? ?
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