Visualizing morphogenesis in transgenic zebrafish embryos using BODIPY TR methyl ester dye as a vital counterstain for GFP.

Green fluorescent protein (GFP) technology is rapidly advancing the study of morphogenesis, by allowing researchers to specifically focus on a subset of labeled cells within the living embryo. However, when imaging GFP-labeled cells using confocal microscopy, it is often essential to simultaneously visualize all of the cells in the embryo using dual-channel fluorescence to provide an embryological context for the cells expressing GFP. Although various counterstains are available, part of their fluorescence overlaps with the GFP emission spectra, making it difficult to clearly identify the cells expressing GFP. In this study, we report that a new fluorophore, BODIPY TR methyl ester dye, serves as a versatile vital counterstain for visualizing the cellular dynamics of morphogenesis within living GFP transgenic zebrafish embryos. The fluorescence of this photostable synthetic dye is spectrally separate from GFP fluorescence, allowing dual-channel, three-dimensional (3D) and four-dimensional (4D) confocal image data sets of living specimens to be easily acquired. These image data sets can be rendered subsequently into uniquely informative 3D and 4D visualizations using computer-assisted visualization software. We discuss a variety of immediate and potential applications of BODIPY TR methyl ester dye as a vital visualization counterstain for GFP in transgenic zebrafish embryos.

The Zebrafish DVD Exchange Project: a bioinformatics initiative.

Scientists who study zebrafish currently have an acute need to increase the rate of visual data exchange within their international community. Although the Internet has provided a revolutionary transformation of information exchange, the Internet is at present unable to serve as a vehicle for the efficient exchange of massive amounts of visual information. Much like an overburdened public water system, the Internet has inherent limits to the services it can provide. It is possible, however, for zebrafishologists to develop and use virtual intranets (such as the approach we outlined in this chapter) to adapt to the growing informatics need of our expanding research community. We need to assess qualitatively the economics of visual bioinformatics in our research community and evaluate the benefit:investment ratio of our collective information-sharing activities. The development of the World Wide Web started in the early 1990s by particle physicists who needed to rapidly exchange visual information within their collaborations. However, because of current limitations in information bandwidth, the World Wide Web cannot be used to easily exchange gigabytes of visual information. The Zebrafish DVD Exchange Project is aimed at by-passing these limitations. Scientists are curiosity-driven tool makers as well as curiosity-driven tool users. We have the capacity to assimilate new tools, as well as to develop new innovations, to serve our collective research needs. As a proactive research community, we need to create new data transfer methodologies (e.g., the Zebrafish DVD Exchange Project) to stay ahead of our bioinformatics needs.