The proposed research deals with the development of new imaging technologies with the use of semi-conductor and metal nanocrystals These new imaging technologies will continually be tested and used for the study of morphogenesis in Xenopus Laevis and other embryos. The primary goal of this project is to fully utilize the optical properties of QD;s for the study of biological processes. The study of morphogenetic movements in particular, is especially dependent on cutting edge imaging technologies, which in large part set the pace of discovery in the field. Our previous work includes the development of the first biocompatible Quantum Dots which were used for linage tracing and real time in vivo imaging of the development of Xenopus embryos. One of the first projects that we would pursue is the use of Near Infra Red QD;s for labeling and imaging of deep tissues within the Xenopus embryo. Current imaging technologies fail to provide visualization of deep tissue movements in vivo restricting our observation to t he superficial layers and thus limiting our understanding. Development of deep tissue imaging will have several other applications, for example in non invasive diagnostic testing. A second objective would be the labeling of proteins with QD;s and investiga ting the effect of the label on a proteins function. We have successfully labeled actin monomers with QD;s and observed actin dynamics in vivo in Xenopus. We are planning to use Akt;s Plekstrin Homology domain labeled with QD;s as a biosensor to monitor th e distribution of active Akt within migrating cells, in order to investigate its role in the migration of mesodermal cells. Preliminary results obtained using PH-GFP fusions suggest that Akt is involved but GFP fusions photobleach quickly and cannot be ima ged in deep migrating tissues. It is expected that this IRG grant will help the principal investigator to establish a new research laboratory at Intercollege and help initiate his research activities.
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