Core D will implement hyperspectral imaging technologies, image analysis approaches, and mathematical modeling to answer questions concerning the time course and spatial spread of cellular signals and subsequent regulation of endothelial function. Core D will support all PROJECTS by providing spectral analysis of the molecular composition of tissues, enabling quantitative imaging of localized intracellular signals and implementing novel image processing, data analysis, and mathematical modeling approaches. Service component: Core D will provide capabilities for next-generation imaging, image analysis, quantitative data extraction, mathematical modeling, and data storage and retrieval to support all projects and cores. In specific, Core D will 1) provide expertise and assistance in implementing next-generation 5-dimensional imaging (x,y,z,t,?) approaches; 2) develop customized analysis approaches for extracting localized signaling information from 5-dimensional image data; 3) assist in creating mathematical models of signaling pathways to aid in hypothesis testing and generation; and 4) manage a central repository for image and modeling data storage that will facilitate cross-project collaboration and data mining. Academic component: The investigators of Core D will continue to develop new technologies for next- generation hyperpsectral, high-speed microscopy. As these microscope systems are implemented, Core D will provide access to and training on the new imaging technologies. During the upcoming cycle, Core D will focus on implementing several high-speed hyperspectral imaging technologies that are currently in the late stages of prototype development. Access to these technologies will enable project investigators to take the next step in studying pulmonary vasculature on a cellular and biosystems level (for example, simultaneous subcellular measurements of Ca2+-cGMP-NO, or assessing cAMP in whole tissue constructs and intravitally). Synergy with projects and scientific cores: Core D will serve as a hub for project synergy by implementing cross-project quantitative imaging, analysis, and modeling approaches and by ensuring that all image and modeling data is archived and maintained in a central database (repository). The quantitative nature of 5D imaging and the ability to access data from experiments across multiple projects will further build synergy. For example, we can quantitatively assess whether Ca2+ influx through T-type channels in response to a 20 second depolarization is sufficient to modulate sAC and AC6 activity at designated subcellular locations and to effect changes in cAMP and endothelial barrier integrity. Thus, the quantitative nature of Core D provides a framework for interweaving imaging and modeling studies performed by all projects.
Core D is the BioImaging and BioTechnology Implementation Core. Core D will implement hyperspectral imaging technologies, image analysis approaches, and mathematical modeling to answer questions concerning the kinetics and spatial spread of cellular signals and regulation of pulmonary endothelial function.
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